Updated on 2022/05/19

写真a

 
TAKEDA, Naoya
 
Affiliation
Faculty of Science and Engineering, School of Advanced Science and Engineering
Job title
Professor
Profile
1995 JSPS Research Fellow (DC1) 1998 JSPS Research Fellow (PD) 2001 Assistant Professor, Institute of Biomedical Engineering and Science, Tokyo Women’s Medical University 2005 Associate Professor, Institute for Biomedical Engineering, Waseda University 2007 Associate Professor, Dept. of Life Sci. and Med. Biosci., School of Adv. Sci. and Eng., Waseda University 2018 Professor, Dept. of Life Sci. and Med. Biosci., School of Adv. Sci. and Eng., Waseda University

Concurrent Post

  • Faculty of Science and Engineering   Graduate School of Advanced Science and Engineering

  • Affiliated organization   Global Education Center

Research Institute

  • 2020
    -
    2022

    理工学術院総合研究所   兼任研究員

Education

  • 2005
    -
    2007

    Waseda University   Graduate School of Asia-Pacific Studies   Business Administration in Technology Management  

  • 1993
    -
    1998

    The University of Tokyo   Graduate School of Engineering   Department of Chemistry and Biotechnology  

  • 1989
    -
    1993

    The University of Tokyo   Faculty of Engineering   Department of Industrial Chemistry  

Degree

  • 東京大学   博士(工学)

  • 早稲田大学   経営学修士(専門職)

  • The University of Tokyo   Doctor of Engineering

  • Waseda University   Master of Business Administration in Technology Management

Research Experience

  • 2018.04
    -
    Now

    Waseda University   Department of Life Science and Medical Bioscience, School of Advanced Science and Engineering   Professor

  • 2014
    -
    Now

    Tokyo Women's Medical University   Graduate School of Nursing   part-time lecturer

  • 2004
    -
    Now

    Tokyo Women's Medical University   Institute of Advanced Biomedical Engineering and Science   part-time lecturer

  • 2020.04
    -
    2022.03

    東京医科歯科大学 生体材料工学研究所 非常勤講師

  • 2007
    -
    2018

    Waseda University   Department of Life Science and Medical Bioscience, School of Advanced Science and Engineering   Associate Professor

  • 2009
    -
    2016

    University of Yamanashi   Integrated Graduate School of Medicine and Engineering   part-time lecturer

  • 2014
    -
    2015

    Tokyo Medical Dental University   Institute of Biomaterials and Bioengineering   part-time lecturer

  • 2005
    -
    2007

    Waseda University   Institute for Biomedical Engineering   Associate Professor

  • 2004
    -
    2005

    Waseda University   Institute for Biomedical Engineering   Lecturer

  • 2001
    -
    2004

    Tokyo Women's Medical University   Department of Neurosurgery   (Concurrent) Assistant Professor

  • 2001
    -
    2004

    Tokyo Women’s Medical University   Institute of Advanced Biomedical Engineering and Science   Assistant Professor

  • 1998
    -
    2001

    Japan Society for the Promotion of Science   Reseach Fellow (PD)

  • 1995
    -
    1998

    Japan Society for the Promotion of Science   Reseach Fellow (DC1)

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Professional Memberships

  •  
     
     

    Japanese Society for Artificial Organs

  •  
     
     

    The Japan Society of Drug Delivery System

  •  
     
     

    The Japan Neuroscience Society

  •  
     
     

    The Japanese Society for Regenerative Medicine

  •  
     
     

    The Society of Polymer Science, Japan

  •  
     
     

    The Chemical Society of Japan

  •  
     
     

    Japanese Society for Biomaterials

  •  
     
     

    THE CELLULOSE SOCIETY OF JAPAN

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Research Areas

  • Polymer materials   Stimuli-responsive Materials, Sensing Materials

  • Structural materials and functional materials

  • Thin film/surface and interfacial physical properties   Nano film

  • Nanobioscience

  • Polymer chemistry

  • Biomaterials   Culture Scaffold, Hydrogel

  • Biomedical engineering   Cell and Tissue Engineering, Regenerative Medicine

▼display all

Research Interests

  • Biomaterials, Functional Polymer Materials, Soft Interface, Cellular and Tissue Engineering, Regenerative Medicine, Bio-MEMS

Papers

  • Self-healing polyurethane elastomers based on charge-transfer interactions for biomedical applications

    Keiichi Imato, Hidekazu Nakajima, Ryota Yamanaka, Naoya Takeda

    Polymer Journal   53 ( 2 ) 355 - 362  2021.02  [Refereed]

    Authorship:Last author, Corresponding author

    DOI

  • Enhanced mechanical properties and cell separation with thermal control of PIPAAm-brushed polymer-blend microfibers

    Kenichi Nagase, Risa Shukuwa, Hironobu Takahashi, Naoya Takeda, Teruo Okano

    Journal of Materials Chemistry B   8 ( 28 ) 6017 - 6026  2020  [Refereed]

    Authorship:Corresponding author

     View Summary

    <p>Thermoresponsive microfibers with enhanced mechanical properties for temperature-modulated cell separation were developed by electrospinning of blending PVBC and PBMA, and by subsequently modifying the microfibers with PIPAAm <italic>via</italic> ATRP.</p>

    DOI

  • Fluorescent supramolecular mechanophores based on charge-transfer interactions

    Keiichi Imato, Ryota Yamanaka, Hidekazu Nakajima, Naoya Takeda

    Chemical Communications   56 ( 57 ) 7937 - 7940  2020  [Refereed]

    Authorship:Last author, Corresponding author

     View Summary

    <p>Supramolecular mechanofluorophores based on charge-transfer interactions between fluorescent electron-rich pyrene and electron-deficient naphthalene diimide(s) are newly developed and show turn-on fluorescence upon application of mechanical forces.</p>

    DOI

  • Cell adhesion control by photoinduced LCST shift of PNIPAAm-based brush scaffolds

    Keiichi Imato, Kazuho Nagata, Rina Watanabe, Naoya Takeda

    Journal of Materials Chemistry B   8 ( 12 ) 2393 - 2399  2020  [Refereed]

    Authorship:Last author, Corresponding author

     View Summary

    <p>Cell adhesion was markedly regulated on spiropyran-containing PNIPAAm-based brush scaffolds by photoinduced LCST shift across a standard culture temperature.</p>

    DOI

  • A stable protocol for the fabrication of transplantable human oral mucosal epithelial cell sheets for clinical application

    Yoshiyuki Kasai, Ryo Takagi, Shinichiro Kobayashi, Toshiyuki Owaki, Naoyuki Yamaguchi, Hiroko Fukuda, Yusuke Sakai, Yoshinori Sumita, Nobuo Kanai, Hajime Isomoto, Kengo Kanetaka, Takeshi Ohki, Izumi Asahina, Kazuhiro Nagai, Kazuhiko Nakao, Naoya Takeda, Teruo Okano, Susumu Eguchi, Masayuki Yamato

    Regenerative Therapy   in press  2019.12  [Refereed]

  • Poly(N‑isopropylacrylamide)-Grafted Polydimethylsiloxane Substrate for Controlling Cell Adhesion and Detachment by Dual Stimulation of Temperature and Mechanical Stress

    Yoshikatsu Akiyama, Miki Matsuyama, Masayuki Yamato, Naoya Takeda, Teruo Okano

    Biomacromolecules   19   4014 - 4022  2018  [Refereed]

    Authorship:Corresponding author

    DOI

  • Photoresponsive fiber scaffolds with a core-sheath nanostructure for regulating cell behaviors

    Kazuho Nagata, Tetsuya Kurebayashi, Keiichi Imato, Naoya Takeda

    Journal of Materials Chemistry B   6 ( 14 ) 2052 - 2056  2018  [Refereed]

    Authorship:Last author, Corresponding author

     View Summary

    Herein, we report the fabrication of photoresponsive three-dimensional (3D) fiber scaffolds for the first time, where photoresponsive polymers are localized on their fiber surfaces of nano thickness, using a simple and practical co-axial (core-sheath) electrospinning technique. Cell adhesion to the 3D scaffolds was regulated by photostimulation.

    DOI

  • Cellular events and behaviors after grafting of stratified squamous epithelial cell sheet onto a hydrated collagen gel

    Yoshiyuki Kasai, Naoya Takeda, Shinichiro Kobayashi, Ryo Takagi, Masayuki Yamato

    FEBS Open Bio   7 ( 5 ) 691 - 704  2017.05  [Refereed]

    Authorship:Corresponding author

     View Summary

    Autologous stratified squamous epithelial cell sheets have been successfully used to treat epithelial defects in tissues such as the cornea and the esophagus. However, the regenerative cellular events occurring in the grafted epithelial cells are unclear in the early stages of wound healing. In this study, we created an in vitro grafting model using cultured normal human epidermal keratinocyte (NHEK) sheets and a type I collagen gel to investigate the cellular processes that occur within the grafted cell sheet. Cultured NHEK cells successfully became a stratified squamous cell sheet resembling epithelial tissue, retained expression of cellular integrins and adhesion proteins, and adhered successfully to a type I collagen gel. After culture on the collagen gel, expression of E-cadherin, and β-catenin decreased in the cells of the basal layer of the grafted cell sheet, resembling events characteristic of a partial epithelial–mesenchymal transition (EMT). These basal cells also induced migration of the cell sheet. Those phenomena are consistent with the essential events that occur in the wound-healing process observed previously in cell studies. Therefore, the epithelial cell sheet grafted onto a type I collagen gel is a suitable model in vitro to study cellular events and behaviors. Furthermore, we also addressed the therapeutic mechanisms by which the epithelial cell sheet promotes wound healing.

    DOI

  • Thermoresponsive polymer-modified microfibers for cell separations

    Kenichi Nagase, Yoichi Sakurada, Satoru Onizuka, Takanori Iwata, Masayuki Yamato, Naoya Takeda, Teruo Okano

    Acta Biomaterialia   53   81 - 92  2017.04  [Refereed]

    Authorship:Corresponding author

     View Summary

    Thermoresponsive polymer-modified microfibers were prepared through electrospinning of poly(4-vinylbenzyl chloride) (PVBC) and subsequent surface-initiated atom transfer radical polymerization for grafting poly(N-isopropylacrylamide) (PIPAAm). Electrospinning conditions were optimized to produce large-diameter (20 μm) PVBC microfibers. The amount of PIPAAm grafted on the microfibers was controlled via the IPAAm monomer concentration. The microfibers exhibited thermally controlled cell separation by selective adhesion of normal human dermal fibroblasts in a mixed cell suspension that also contained human umbilical vein endothelial cells. In addition, adipose-derived stem cells (ADSCs) exhibited thermally modulated cell adhesion and detachment, while adhesion of other ADSC-related cells was low. Thus, ADSCs could be separated from a mixture of adipose tissue-derived cells simply by changing the temperature. Overall, the PIPAAm-modified microfibers are potentially applicable as temperature-modulated cell separation materials. Statement of Significance Thermoresponsive poly(N-isopropylacrylamide) (PIPAAm) polymer-modified poly(4-vinylbenzyl chloride) (PVBC) microfibers were prepared via electrospinning of PVBC, followed by surface-initiated ATRP. They formed effective thermally-modulated cell separation materials with large surface areas. Cells adhered and extended along the modified microfibers
    this was not observed on previously reported PIPAAm-modified flat substrates. The cellular adhesion enabled separation of fibroblast cells, as well as that of adipose-derived mesenchymal stem cells, from mixtures of similar cells. Thus, the temperature-controlled thermoresponsive microfibers would be potentially useful as cell separation materials.

    DOI PubMed

  • Micro/nano-imprinted substrates grafted with a thermoresponsive polymer for thermally modulated cell separation

    Kenichi Nagase, Risa Shukuwa, Takahiro Onuma, Masayuki Yamato, Naoya Takeda, Teruo Okano

    Journal of Materials Chemistry B   5 ( 30 ) 5924 - 5930  2017  [Refereed]

    Authorship:Corresponding author

     View Summary

    There is a great demand for effective cell separation techniques that do not require the labeling of cell surfaces for applications in cell transplantation therapy and cell analysis. In the present study, we prepared thermoresponsive convex or concave substrates with circular hole, cylindrical pillar, and line patterns of various sizes as thermally modulated cell separation materials through the combination of thermal nano-imprinted lithography and subsequent surface-initiated atom transfer radical polymerization of poly(N-isopropylacrylamide). Three types of human cells, human umbilical vein endothelial cells, normal human dermal fibroblasts, and human skeletal muscle myoblast cells, which are commonly used in cardiovascular tissue engineering, were employed in this study. All three cell types could adhere to the prepared thermoresponsive micro- or nano-imprinted substrates at 37 °C and detached at 20 °C. The specific cell adhesion and detachment properties were different for each cell type, and they could be altered simply by changing the pattern shapes and sizes of the surface. In particular, large differences between the three cell types were obtained on the 2 μm hole pattern. Using this difference in cell adhesion properties, thermally modulated cell separation application was achieved by successively incubating at 37 °C and 20 °C. Thus, our thermoresponsive micro/nano-imprinted substrates can be utilized as cooperative cell separating materials by combining appropriate convex or concave patterns and mild temperature changes.

    DOI

  • A Photoresponsive Soft Interface Reversibly Controls Wettability and Cell Adhesion by Conformational Changes in a Spiropyran-Conjugated Amphiphilic Block Copolymer

    Di He, Yoshinori Arisaka, Kenichi Masuda, Mitsuya Yamamoto, Naoya Takeda

    Acta Biomaterialia   51   101 - 111  2017  [Refereed]

    Authorship:Last author, Corresponding author

     View Summary

    © 2017 Acta Materialia Inc. The functionalities of soft interfaces including cell adhesion can be enhanced by dynamic conversion of polymer properties and movement via external stimuli. Light is a superior stimulus, and various surfaces modified with photoreactive molecules have been prepared. However, in most of these studies, the surface properties are irreversibly changed due to photo-degradation, and reversible adhesion and collection of cells is not feasible. In this study, we developed a photoresponsive polymer soft interface that was able to spatiotemporally control wettability, cell adhesion, and detachment in a reversible manner. Spiropyran molecules were introduced into the hydrophobic block of an amphiphilic diblock copolymer consisting of poly(methyl methacrylate) and polyethylene glycol, and the monomer unit numbers of these components were optimized. The copolymer was immobilized on a glass substrate as a nanofilm. With alternating irradiation using UV and visible light, the surface exhibited reversible changes in hydrophobicity and hydrophilicity, and the direction of change was opposite to the polarity change in photo-isomerization of spiropyran. We also achieved p

    DOI

  • Photo-induced in situ crosslinking of polymer brushes with dimethyl maleimide moieties for dynamically stimulating stem cell differentiation

    Yoshinori Arisaka, Yuka Nishijima, Shin-Ichi Yusa, Naoya Takeda

    Journal of Biomaterials Science, Polymer Edition   27 ( 13 ) 1331 - 1340  2016.09  [Refereed]

    Authorship:Last author, Corresponding author

     View Summary

    We designed photo-crosslinkable polymer brushes with dimethylmaleimide moieties, in order to demonstrate dynamic stimulation of cell differentiation in mesenchymal stem cells (MSCs). The polymer brushes were synthesized by surface-initiated reversible addition fragmentation chain transfer polymerization using dimethylmaleimide ethyl methacrylate and methyl methacrylate on a chain transfer agent-immobilized glass surface. The polymer brushes were crosslinked by photodimerization of the dimethylmaleimide moieties within polymer chains with stem cells present on the surface. In order to evaluate the effects of in situ photo-induced crosslinking of the polymer brushes on gene expression of stem cells, human bone marrow MSCs were cultured under static and dynamic culture conditions for 7 days. Expression of the osteocalcin (Ocn) gene in MSCs was used as an indicator of osteoblast differentiation under dynamic culture conditions. Structural conversion from non-crosslinked polymer brushes to crosslinked polymer brushes increased the expression of Ocn by 1.4-fold in the presence of adhered cells, compared with non-crosslinked polymer brushes under static culture conditions. These results suggest that MSCs recognized surface conversion from non-crosslinked to crosslinked structures, which resulted in altered differentiation lineages. Therefore, photo-crosslinkable surfaces with dimethyl maleimide moieties are potential novel materials for dynamically stimulating MSC differentiation.

    DOI

  • Brush biopsy of human oral mucosal epithelial cells as a quality control of the cell source for fabrication of transplantable epithelial cell sheets for regenerative medicine.

    Yoshiyuki Kasai, Hiroaki Sugiyama, Ryo Takagi, Makoto Kondo, Toshiyuki Owaki, Hideo Namiki, Teruo Okano, Naoya Takeda, Masayuki Yamato

    Regenerative therapy   4   71 - 77  2016.06  [Refereed]  [International journal]

     View Summary

    Autologous oral mucosal epithelial cell sheets have been used for treating epithelial defects such as cornea and esophagus. The cell source of patients' oral mucosal epithelial cell sheet should be examined in normality because it has individual difference. In this study, oral mucosal epithelial cells were less invasively collected by brush biopsy from the buccal, gingival, labial, and palate mucosa of four healthy volunteer donors without anesthesia, and analyzed the keratin expressions by western blotting and the obtained results were compared with those by immunohistochemistry of each of the native tissues. All of the oral mucosal epithelial cells expressed keratin 4 (K4) and K13, which were mucosal stratified squamous epithelial cell markers. K1 and K10, keratinized epithelial cell markers, were also detected in keratinized tissues such as gingival and palate mucosa. The markers of epithelial basal cells such as p63 and K15 were not detected by brush biopsy-western blotting. Although this method does not include basal layers of oral mucosa, protein expressions of upper layer of lesion area are different from normal. Therefore, brush biopsy-western blotting was extremely less invasive and would contribute to quality control of the fabrication of autologous oral mucosal epithelial cell sheets.

    DOI PubMed

  • In situ cross-linked electrospun fiber scaffold of collagen for fabricating cell-dense muscle tissue

    Naoya Takeda, Kenichi Tamura, Ryo Mineguchi, Yumiko Ishikawa, Yuji Haraguchi, Tatsuya Shimizu, Yusuke Hara

    Journal of Artificial Organs   19 ( 2 ) 141 - 148  2016.06  [Refereed]

    Authorship:Lead author, Corresponding author

     View Summary

    Engineered muscle tissues used as transplant tissues in regenerative medicine should have a three-dimensional and cell-dense structure like native tissue. For fabricating a 3D cell-dense muscle tissue from myoblasts, we proposed the electrospun type I collagen microfiber scaffold of the string-shape like a harp. The microfibers were oriented in the same direction to allow the myoblasts to align, and were strung at low density with micrometer intervals to create space for the cells to occupy. To realize this shape of the scaffold, we employed in situ cross-linking during electrospinning process for the first time to collagen fibers. The collagen microfibers in situ cross-linked with glutaraldehyde stably existed in the aqueous media and completely retained the original shape to save the spaces between the fibers for over 14 days. On the contrary, the conventional cross-linking method by exposure to a glutaraldehyde aqueous solution vapor partially dissolved and damaged the fiber to lose a low-density shape of the scaffold. Myoblasts could penetrate into the interior of the in situ cross-linked string-shaped scaffold and form the cell-dense muscle tissues. Histochemical analysis showed the total area occupied by the cells in the cross section of the tissue was approximately 73 %. Furthermore, the resulting muscle tissue fabricated from primary myoblasts showed typical sarcomeric cross-striations and the entire tissue continuously pulsated by autonomous contraction. Together with the in situ cross-linking, the string-shaped scaffold provides an efficient methodology to fabricate a cell-dense 3D muscle tissue, which could be applied in regenerative medicine in future.

    DOI PubMed

  • Stem cell separation using thermoresponsive copolymer brushes having cationic charge

    Kenichi Nagase, Yuri Hatakeyama, Tatsuya Shimizu, Katsuhisa Matsuura, Masayuki Yamato, Naoya Takeda, Teruo Okano

    2015 International Symposium on Micro-NanoMechatronics and Human Science, MHS 2015    2016.03

     View Summary

    For developing new stem cell separation tool, we have prepared thermoresponsive cationic copolymer brush, poly(N-isopropylacrylamide-co-N, N-dimethylaminopropylacrylamide-co-N-tert-butylacrylamide (IPAAm-co-DMAPAAm-co-tBAAm) on glass substrates through surface-initiated atom transfer radical copolymerization (ATRP). Using the prepared thermoresponsive cationic copolymer brush, mesenchymal stem cell separation was performed simply by changing temperature.

    DOI

  • Self-Driven Perfusion Culture System Using a Paper-Based Double-Layered Scaffold

    Ai Ozaki, Yoshinori Arisaka, Naoya Takeda

    Biofabrication   8   035010  2016  [Refereed]

    Authorship:Last author, Corresponding author

     View Summary

    © 2016 IOP Publishing Ltd.Shear stress caused by fluid flow is known to promote tissue development from cells in vivo. Therefore, perfusion cultures have been studied to investigate the mechanisms involved and to fabricate engineered tissues in vitro, particularly those that include blood vessels. Microfluidic devices, which function with fine machinery of chambers and microsyringes for fluid flow and have small culture areas, are conventionally used for perfusion culture. In contrast, we have developed a self-driven perfusion culture system by using a paper-based double-layered scaffold as the fundamental component. Gelatin microfibers were electrospun onto a paper material to prepare the scaffold system, in which the constant perfusion of the medium and the scaffold for cell adhesion/proliferation were functionally divided into a paper and a gelatin microfiber layer, respectively. By applying both the capillary action and siphon phenomenon of the paper-based scaffold, which bridged two medium chambers at different height levels, a self-driven medium flow was achieved and the flow rate was also stable, constant, and quantitatively controllable. Moreover, the culture area was enla

    DOI

  • Enhanced Wettability Changes by Synergistic Effect of Micro/Nanoimprinted Substrates and Grafted Thermoresponsive Polymer Brushes

    Kenichi Nagase, Takahiro Onuma, Masayuki Yamato, Naoya Takeda, Teruo Okano

    Macromolecular Rapid Communications   36 ( 22 ) 1965 - 1970  2015.11  [Refereed]

    Authorship:Corresponding author

     View Summary

    Thermoresponsive polymer brushes are grafted on micro/nanostructured polymer substrates as new intelligent interfaces that synergistically enhance wettability changes in response to external temperature stimuli. Thermoplastic poly(styrene-co-4-vinylbenzyl chloride) [P(St-co-VBC)] is synthesized using radical polymerization and spin-coated on a glass substrate. Micro/nanopillar and hole patterns are imprinted on the P(St-co-VBC) layer using thermal nanoimprint lithography. Poly(N-isopropylacrylamide) (PIPAAm) brushes are grafted on the micro/nanostructured P(St-co-VBC) layer through surface-initiated atom-transfer radical polymerization using 4-vinylbenzyl chloride as the initiator. The imprinted micro/nanostructures and grafted PIPAAm brush chain lengths affect the surface wettability. Combinations of nanopillars or nanoholes (diameter 500 nm) and longer PIPAAm brushes enhance hydrophobic/hydrophilic changes in response to temperature changes, compared with the flat substrate. The thermoresponsive hydrophobic/hydrophilic transition is synergistically enhanced by the nanostructured surface changing from Cassie-Baxter to Wenzel states. This PIPAAm-brush-modified micro/nanostructured P(St-co-VBC) is a new intelligent interface that effectively changes wettability in response to external temperature changes. Novel thermoresponsive interfaces showing significant wettability changes are fabricated by nanoimprinting a poly(styrene-co-4-vinylbenzyl chloride) layer and subsequent surface-initiated atom transfer radical polymerization of poly(N-isopropylacrylamide) (IPAAm). Combinations of nanopillars or nanoholes and longer PIPAAm brushes enhance hydrophobic/hydrophilic changes in response to temperature changes, compared with a flat substrate.

    DOI PubMed

  • How to prevent contamination with Candida albicans during the fabrication of transplantable oral mucosal epithelial cell sheets.

    Ryo Takagi, Shinichiro Kobayashi, Masayuki Yamato, Toshiyuki Owaki, Yoshiyuki Kasai, Takahiro Hosoi, Yusuke Sakai, Kengo Kanetaka, Tokutaro Minamizato, Asuka Minematsu, Makoto Kondo, Nobuo Kanai, Naoyuki Yamaguchi, Kazuhiro Nagai, Yasushi Miyazaki, Naoya Takeda, Fumio Fukai, Izumi Asahina, Taiga Miyazaki, Shigeru Kohno, Masakazu Yamamoto, Kazuhiko Nakao, Susumu Eguchi, Teruo Okano

    Regenerative therapy   1   1 - 4  2015.06  [Refereed]  [International journal]

     View Summary

    We have utilized patients' own oral mucosa as a cell source for the fabrication of transplantable epithelial cell sheets to treat limbal stem cell deficiency and mucosal defects after endoscopic submucosal dissection of esophageal cancer. Because there are abundant microbiotas in the human oral cavity, the oral mucosa was sterilized and 40 μg/mL gentamicin and 0.27 μg/mL amphotericin B were added to the culture medium in our protocol. Although an oral surgeon carefully checked each patient's oral cavity and although candidiasis was not observed before taking the biopsy, contamination with Candida albicans (C. albicans) was detected in the conditioned medium during cell sheet fabrication. After adding 1 μg/mL amphotericin B to the transportation medium during transport from Nagasaki University Hospital to Tokyo Women's Medical University, which are 1200 km apart, no proliferation of C. albicans was observed. These results indicated that the supplementation of transportation medium with antimycotics would be useful for preventing contamination with C. albicans derived from the oral mucosa without hampering cell proliferation.

    DOI PubMed

  • One core-five sheaths coaxial flow formation using multilayer stacked flow focusing structure

    D. H. Yoon, J. Ito, N. Takeda, T. Sekiguchi, S. Shoji

    Proceedings of the IEEE International Conference on Micro Electro Mechanical Systems (MEMS)   2015- ( February ) 539 - 542  2015.02  [Refereed]

     View Summary

    This research presents efficient coaxial sheath flow formation using a three-dimensional PDMS device. In the device, one core and five sheaths are simply formed with low diffusion between different samples. Stacking and alignment of six PDMS layers of same structure allowed for fabrication of the proposed 3D device. Only one point alignment of center channel for the sheath provides free from misalignment that was a critical problem of multi-stacked PDMS device. Furthermore, the number of samples is infinitely expandable by increase in the number of stacking layers. The coaxial sheath flow is useful for biological fiber formation which requires multilayer of different materials such as artificial blood vessels or muscles.

    DOI

  • Highly controllable three-dimensional sheath flow device for fabrication of artificial capillary vessels

    J. Ito, R. Sekine, D. H. Yoon, Y. Nakamura, H. Oku, H. Nansai, T. Chikasawa, T. Goto, T. Sekiguchi, N. Takeda, S. Shoji

    Proceedings of the IEEE International Conference on Micro Electro Mechanical Systems (MEMS)   2015- ( February ) 480 - 483  2015.02  [Refereed]

     View Summary

    This paper reports a highly controllable three-dimensional (3D) sheath flow device for fabrication of artificial capillary vessels. Three step sheath injection type 3D flow device which realizes wide core and sheath structure variations by simply flow rate control was applied to fabricate double-layer coaxial Core-Sheath microfibers applicable for long micro capillary vessels by aligning and cultivating vascular endothelial cells. As a result, about a 3-centimeter-long microfiber which has the vascular endothelial cells fused mutually in the center was successfully formed after three days culture.

    DOI

  • Thermoresponsive cationic copolymer brushes for mesenchymal stem cell separation

    Kenichi Nagase, Yuri Hatakeyama, Tatsuya Shimizu, Katsuhisa Matsuura, Masayuki Yamato, Naoya Takeda, Teruo Okano

    Biomacromolecules   16 ( 2 ) 532 - 540  2015.02  [Refereed]

     View Summary

    Thermoresponsive, cationic, copolymer brushes poly(N-isopropylacrylamide(IPAAm)-co-N,N-dimethylaminopropylacrylamide-co-N-tert-butylacrylamide(tBAAm)) and poly(IPAAm-co-3-acrylamidopropyl trimethylammonium chloride-co-tBAAm) were prepared on glass substrates through surface-initiated atom transfer radical polymerization. Prepared copolymer brushes were investigated as thermally modulated cell separation materials. Densely packed cationic copolymer brushes were formed on the glass substrates, and the positive charge density was modulated by controlling the composition of cationic moieties and species. During observation of cell adhesion and detachment properties on copolymer brushes, human bone marrow mesenchymal stem cells (hbmMSC) exhibited thermally modulated cell adhesion and detachment, while other bone-marrow-derived cells did not adhere. Using these properties, hbmMSC could be purified from mixtures of human bone-marrow-derived cells simply by changing the external temperature. Therefore, the prepared cationic copolymer brush is useful for separation of hbmMSC.

    DOI PubMed

  • Hydrophobized Thermoresponsive Copolymer Brushes for Cell Separation by Multistep Temperature Change

    Kenichi Nagase, Yuri Hatakeyama, Tatsuya Shimizu, Katsuhisa Matsuura, Masayuki Yamato, Naoya Takeda, Teruo Okano

    Biomacromolecules   14   3423 - 3433  2013  [Refereed]

    DOI

  • Spatiotemporally controlled navigation of neurite outgrowth in sequential steps on the dynamically photo-patternable surface

    Yoshikuni Edagawa, Jun Nakanishi, Kazuo Yamaguchi, Naoya Takeda

    Colloids and Surfaces B: Biointerfaces   99   20 - 26  2012.11  [Refereed]

    Authorship:Last author, Corresponding author

     View Summary

    A great number of the neurites interconnect neuronal cells in a brain to form the complicate neural circuits, whose structures are dynamically changed with changing the numbers and destinations of the neurites. Fabricating a model of neural network in vitro is one of the promising methods to precisely assay the signal transmission and processing within the circuit as well as to examine behaviors of individual cells. In this study, aiming to fabricate the dynamically alterable neural network in vitro, the chemically modified surface with the photo-reactive self-assembled monolayer was applied to navigate the neurite outgrowth activities of differentiated PC12 cell in the spatially and temporally controlled manner. Numbers of the cell soma were effectively adhered and simultaneously arrayed according to the 25 μm square patterns, which were easily fabricated with a single shot of the 365-nm ultraviolet (UV) irradiation and pre-coated with the extracellular matrix (ECM) protein. Narrow neurites were successively guided along the 5 μm line patterns drawn on the surface by stepwise irradiation of the UV light in the intended designs and at appropriate timing. Sprouting number, elongating direction, bending, branching, and formation of autapse-like structure were controllable. The rate of neurite elongation was dependent on the ECM species, that were pre-coated beneath the cell soma, suggesting the ECM stimulated the basal side of the cell soma and affected the outgrowth process of the neurite. Navigation of the neurite elongation along the microline pattern for a primary rat brain cortex neuron was also achieved. © 2011 Elsevier B.V.

    DOI PubMed

  • Dynamically cell separating thermo-functional biointerfaces with densely packed polymer brushes

    Kenichi Nagase, Ayaka Kimura, Tatsuya Shimizu, Katsuhisa Matsuura, Masayuki Yamato, Naoya Takeda, Teruo Okano

    Journal of Materials Chemistry   22 ( 37 ) 19514 - 19522  2012.10  [Refereed]

     View Summary

    Poly(N-isopropylacrylamide) (PIPAAm) brush grafted glass surfaces with various brush lengths were prepared as cell separating intelligent interfaces through a surface-initiated atom transfer radical polymerization (ATRP) with a CuCl-Me 6TREN catalytic system and α-chloro-p-xylene as a free initiator in 2-propanol at 25 °C for 16 h. Characterization of the prepared surface was performed by X-ray photoelectron spectroscopy (XPS), attenuated total reflection Fourier transform infrared (ATR/FT-IR) spectroscopy, and gel permeation chromatography (GPC) measurement of PIPAAm in ATRP reaction solution for estimating the brush length. Phase transition behavior of PIPAAm in four cell culture media was also investigated by measuring the temperature-dependent turbidities. Prepared PIPAAm brush surfaces as cell separating intelligent interfaces were characterized by observing the adhesion and detachment behavior of four types of human cells: human umbilical vein endothelial cells (HUVECs), normal human dermal fibroblasts (NHDFs), human aortic smooth muscle cells (SMCs), and human skeletal muscle myoblast cells (HSMMs). The PIPAAm brush surface with a moderate brush length exhibited a proper cell adhesion and detachment behavior, while short-brush-surfaces scarcely detached cells and long-brush-surfaces scarcely adhered cells. The PIPAAm brush with a moderate brush length exhibited different cell detachment rates among individual cell types. Utilizing the different cell detachment properties, a mixture of green fluorescent protein (GFP) expressing HUVEC (GFP-HUVEC) and HSMM was separated. After being allowed to adhere on the surfaces at 37 °C for 24 h, the adhered cells on the surfaces were incubated at 20 °C. In the initial period of incubation at 20 °C, the GFP-HUVEC was released from the surface due to its prompt detachment property, and, in the subsequent period of incubation, HSMMs gradually detached themselves from the surface. These results indicated that a precisely designed PIPAAm brush functioned as an intelligent cell separating interface by utilizing the intrinsic cell detachment properties of individual cells. © 2012 The Royal Society of Chemistry.

    DOI

  • Size-influence in Cell Community Against Apoptotic Impact

    Ryo Harigai, Kentaro Matsuki, Naoya Takeda, Naonobu Shimamoto, Yoshikuni Edagawa

    JOURNAL OF PHARMACOLOGICAL SCIENCES   115   224P  2011

  • Plasmid DNA-encapsulating liposomes: Effect of a spacer between the cationic head group and hydrophobic moieties of the lipids on gene expression efficiency

    Yosuke Obata, Shunsuke Saito, Naoya Takeda, Shinji Takeoka

    Biochimica et Biophysica Acta - Biomembranes   1788 ( 5 ) 1148 - 1158  2009.05  [Refereed]

     View Summary

    We have synthesized a series of cationic amino acid-based lipids having a spacer between the cationic head group and hydrophobic moieties and examined the influence of the spacer on a liposome gene delivery system. As a comparable spacer, a hydrophobic spacer with a hydrocarbon chain composed of 0, 3, 5, 7, or 11 carbons, and a hydrophilic spacer with an oxyethylene chain (10 carbon and 3 oxygen molecules) were investigated. Plasmid DNA (pDNA)-encapsulating liposomes were prepared by mixing an ethanol solution of the lipids with an aqueous solution of pDNA. The zeta potentials and cellular uptake efficiency of the cationic liposomes containing each synthetic lipid were almost equivalent. However, the cationic lipids with the hydrophobic spacer were subject to fuse with biomembrane-mimicking liposomes. 1,5-Dihexadecyl-N-lysyl-N-heptyl-l-glutamate, having a seven carbon atom spacer, exhibited the highest fusogenic potential among the synthetic lipids. Increased fusion potential correlated with enhanced gene expression efficiency. By contrast, an oxyethylene chain spacer showed low gene expression efficiency. We conclude that a hydrophobic spacer between the cationic head group and hydrophobic moieties is a key component for improving pDNA delivery. © 2009 Elsevier B.V. All rights reserved.

    DOI PubMed

  • Realization, characterization and functionalization of lipidic wrapped carbon nanotubes

    Gianni Ciofani, Yosuke Obata, Izumi Sato, Yosuke Okamura, Vittoria Raffa, Arianna Menciassi, Paolo Dario, Naoya Takeda, Shinji Takeoka

    Journal of Nanoparticle Research   11 ( 2 ) 477 - 484  2009.02  [Refereed]

     View Summary

    Mass-produced carbon nanotubes (CNTs) are strongly aggregated and highly hydrophobic, and processes to make them water soluble are required for biological applications. Both covalent and non-covalent strategies are pursued for obtaining stable, highly concentrated CNT aqueous dispersions. Covalent functionalization has the great disadvantage of producing an irreversible chemical modification of nanotubes, thus alterating their mechanical, chemical and electric properties. On the other hand, non-covalent functionalization is often obtained by employing surfactants that sensibly affect cell viability. Moreover, derivatization with biological moieties is often impossible through non-covalent CNT dispersion. This paper proposes a non-covalent dispersion of multi-wall CNT based on a lipidic mixture that can guarantee high concentration and high stability as well as high cytocompatibility. Moreover, CNTs wrapped with a lipid membrane are realized to demonstrate that the proposed CNTs can be functionalised with a dodecapeptide that specifically recognizes activated platelets without chemical modification of the nanotube itself. © 2008 Springer Science+Business Media B.V.

    DOI

  • Development of a poly-dimethylsiloxane microfluidic device for single cell isolation and incubation

    Yoshinori Yamaguchi, Takahiro Arakawa, Naoya Takeda, Yoshikuni Edagawa, Shuichi Shoji

    Sensors and Actuators, B: Chemical   136 ( 2 ) 555 - 561  2009  [Refereed]

     View Summary

    Although single cell manipulation has been developed for precise understanding of cell biology, it is still difficult to handle single cells because of their morphology including the size variation and the flexibility. We developed a single cell manipulation device featuring microchannels and micropockets for single cell capture and cultivation. Single cells were captured noninvasively and sequentially, and each cell was repeatedly sub-cultured to four generations. The single cell manipulation devicewas microfabricated with two main parallel channels allowing the cell suspension and the carrier flow to be injected separately. Those channels, that are main channel and buffer channel, were connected with a narrow (3μm) drain channel, and single cell capture pockets were placed at the point where the main channels and the drain channel connected. A gentle flow was produced in the drain channel because of the difference in the flow rate between the main channel and buffer channel, realized the individual single cell isolation in the capture pocket. When a single cell was captured in a single cell pocket, the captured cell capped the drain channel and caused the other cells that were flowing through the channel to go to the next capture pockets. The ratio between the cells thatwere captured and the cells that passed through the main channel was about 70%. The captured singe cellwas cultured successively in the same pocket, and the cells divided into four cells. The doubling times of two cells that grew in the first cell division were slightly different (10 min). These fundamental single cell manipulations were carried out mainly by controlling the flow rate, essentially the pressure on each channel. Occasionally the manipulations also were carried out by changing the shapes and the sizes of the micropockets in the microfluidic device. Since this device was used successfully for the noninvasive isolation and long-term cultivation of single cells, it can contribute to various biological analyses, such as biopsy, noninvasive bioanalysis, and the morphology of single cells. © 2008 Elsevier B.V. All rights reserved.

    DOI

  • Active species for Ce(IV)-induced hydrolysis of phosphodiester linkage in cAMP and DNA

    Jun Sumaoka, Kenichiro Furuki, Yuki Kojima, Masahiko Shibata, Kimihiko Hirao, Naoya Takeda, Makoto Komiyama

    Nucleosides, Nucleotides and Nucleic Acids   25 ( 4-6 ) 523 - 538  2006.06  [Refereed]

     View Summary

    The hydrolysis of cyclic adenosine 3′,5′-monophosphate and 2′-deoxythymidylyl(3′-5′)2′-deoxythymidine by Ce(NH 4)2(NO3)6 was kinetically studied. The rate of hydrolysis was fairly proportional to the concentration of [Ce 2IV (OH)4]4+, showing that this is the catalytically active species. According to quantum-chemical calculation, the two Ce(IV) ions in this [Ce2IV (OH)4] 4+ cluster are bridged by two OH residues. Upon the complex formation with H2PO4- (a model compound for the phosphodiesters), these two Ce(IV) ions bind the two oxygen atoms of the substrate and enhance the electrophilicity of the phosphorus atom. The catalytic mechanism of Ce(IV)-induced hydrolysis of phosphodiesters has been proposed on the basis these results. Copyright © Taylor &amp
    Francis Group, LLC.

    DOI PubMed

  • Temperature-responsive polymeric carriers incorporating hydrophobic monomers for effective transfection in small doses

    Naoya Takeda, Emiko Nakamura, Masayuki Yokoyama, Teruo Okano

    Journal of Controlled Release   95 ( 2 ) 343 - 355  2004.03  [Refereed]

    Authorship:Lead author

     View Summary

    A series of thermoresponsive ternary random copolymers, poly[N-isopropylacrylamide (PIPAAm)-co-(dimethylamino)ethylmethacrylate (DMAEMA)-co-butylmethacrylate (BMA)], was synthesized and their in vitro gene transfection efficiency in cell culture was evaluated. A control copolymer containing 20 mol% DMAEMA units, IP-20D (mole ratio of IPAAm/DMAEMA/BMA=80/20/0 in feed, no BMA units) was inert in transfection. In contrast, copolymer IP-20D-10B (IPAAm/DMAEMA/BMA=70/20/10 in feed) effectively transfected plasmid DNA into COS-1 cell cultures even under small dosing conditions of 0.1 μg of plasmid DNA per well in a 96-well plate, suggesting that incorporation of the appropriate amount of hydrophobic unit is crucial to transfection efficiency. Gene expression was much more significant when transfected by the IP-20D-10B carrier in comparison with control homopolymer poly-DMAEMA, and almost equal to that of the highly competent lipid carrier, LipofectAMINE PLUS™. Furthermore, the transfection efficiency of IP-20D-10B is altered in a thermally responsive manner. By temporarily lowering the cell culture incubation temperature to 20°C in the posttransfection period, gene expression doubled over that for incubation temperature at 37°C. The DNA EtBr intercalation assay suggested that DNA affinity for IP-20D-10B is decreased by lowering incubation temperature, implying that the thermally regulated gene expression could provide more efficient DNA release from the polymeric carrier. © 2004 Elsevier B.V. All rights reserved.

    DOI PubMed

  • Kinetic and theoretical studies on the mechanism of alkaline hydrolysis of DNA

    Naoya Takeda, Masahiko Shibata, Nobuo Tajima, Kimihiko Hirao, Makoto Komiyama

    Journal of Organic Chemistry   65 ( 14 ) 4391 - 4396  2000.07  [Refereed]

    Authorship:Lead author

     View Summary

    The reaction mechanism of alkaline hydrolysis of DNA has been investigated by kinetic analysis and density-functional-theory calculation. The rates of hydrolysis of thymidine 3'-monophosphate esters (including thymidylyl(3'-5')thymidine (Tp-OT)) monotonically decrease as the leaving groups get poorer. According to the theoretical calculation in which the solvent effects are incorporated, no intermediate is formed in the course of the reaction. In the alkaline hydrolysis of the activated Tp-OT analogues having good leaving groups, the 3',5'-cyclic monophosphate of thymidine is concurrently formed through the intramolecular attack by the 5'-alkoxide ion. In the hydrolysis of the native dinucleotide, however, this side reaction does not occur, since the transition state leading to the departure of its poor leaving group cannot be formed due to conformational restraint. These arguments are supported by the theoretical analysis on the hydrolysis of both dimethyl phosphate and its O(bridging)→ substituted analogue.

    DOI PubMed

  • Hydrolysis of DNA and RNA by lanthanide ions: Mechanistic studies leading to new applications

    Makoto Komiyama, Naoya Takeda, Hidemi Shigekawa

    Chemical Communications   ( 16 ) 1443 - 1451  1999.08  [Refereed]

     View Summary

    A few years ago, the remarkable catalytic activity of lanthanide ions for the hydrolysis of nucleic acids was discovered. With Ce(IV), DNA was hydrolysed under physiological conditions. For RNA hydrolysis, the last three lanthanide ions (Tm(III), Yb(III), and Lu(III)) are superb. Furthermore, artificial restriction enzymes for site-selective scission of DNA and RNA, essential tools for the future biotechnology, have been prepared by using the lanthanide complexes. The present article emphasizes the mechanistic aspects of the catalyses of these metal ions. Both DNA hydrolysis and RNA hydrolysis involve the cooperation of acid catalysis (by metal ion and/or metal-bound water) and base catalysis (by metal-bound hydroxide). The magnitudes of contributions of these catalyses, as well as the positions where they work, are primarily governed by the relative height of the energy-barrier for the formation of the pentacoordinated intermediate and that for its breakdown. The following conclusions have been obtained on the basis of various kinetic and spectroscopic evidence: (1) for the hydrolysis of both DNA and RNA, the catalytically active species are dinuclear hydroxo-clusters, (2) Ce(IV) enormously activates DNA and promotes the formation of the pentacoordinated intermediate, and (3) the catalysis for RNA hydrolysis is mainly ascribed to the promotion of breakdown of the pentacoordinated intermediate.

    DOI

  • DNA Hydrolysis by Ce(IV) Involves the Cooperation of Two Metal Ions

    Jun Sumaoka, Naoya Takeda, Yusuke Okada, Hideyuki Takahashi, Hidemi Shigekawa

    Nucleic Acids, Symp. Ser.   39   137 - 138  1998  [Refereed]

  • Non-enzymatic Hydrolyses of Aryl Esters of Thymidine 3’-Monophosphate as Probes for the Rate-determining Step in DNA Hydrolysis

    Naoya Takeda, Yusuke Okada, Morio Yashiro, Makoto Komiyama

    Nucleic Acids, Symp. Ser.   37   263 - 264  1997  [Refereed]

    Authorship:Lead author

  • Unprecedentedly fast DNA hydrolysis by the synergism of the cerium(IV)-praseodymium(III) and the cerium(IV)-neodymium(III) combinations

    Naoya Takeda, Takamitsu Imai, Makoto Irisawa, Jun Sumaoka, Mono Yashiro, Hidemi Shigekawa, Makoto Komiyama

    Chemistry Letters   25 ( 8 ) 599 - 600  1996  [Refereed]

    Authorship:Lead author

     View Summary

    The most active catalysts for DNA hydrolysis so far have been obtained by using the synergetic cooperation of the cerium(IV) ion with the praseodymium(III) ion. The pseudo first-order rate constant for the hydrolysis of thymidylyl(3'-5')thymidine by the 2:1 combination is 10 times as great as that by the cerium(IV) ion.

    DOI

  • Synergism of Ce(IV) and Pr(III) for the Fastest Non-enzymatic DNA Hydrolysis Ever

    Naoya Takeda, M. Irisawa, Takamitsu Imai, Morio Yashiro, Makoto Komiyama

    Nucleic Acids, Symp. Ser.   34   207 - 208  1995  [Refereed]

    Authorship:Lead author

  • Efficient and Oxygen-Independent Hydrolysis of Single-Stranded DNA by Cerium(IV) Ion

    Makoto Komiyama, Naoya Takeda, Yota Takahashi, Hiroshi Uchida, Tetsuro Shiiba, Teruyuki Kodama, Morio Yashiro

    J. Chem. Soc., Perkin Trans. 2     269 - 274  1995  [Refereed]

    DOI

  • Synergetic catalysis by two non-lanthanide metal ions for hydrolysis of diribonucleotides

    Makoto Irisawa, Naoya Takeda, Makoto Komiyama

    Journal of the Chemical Society, Chemical Communications   ( 12 ) 1221 - 1222  1995  [Refereed]

     View Summary

    Adenylyl(3′-5′)adenosine and uridylyl(3′-5′)uridine are efficiently hydrolysed at pH 7 by bimetallic cooperation of zinc(ii) with tin(IV), indium(iii), iron(iii) or aluminium(iii).

    DOI

  • DNA Hydrolysis by Cerium(IV) Does Not Involve either Molecular Oxygen or Hydrogen Peroxide

    Makoto Komiyama, Tetsuro Shiiba, Teruyuki Kodama, Naoya Takeda, Jun Sumaoka, Morio Yashiro

    Chem. Lett.     1025 - 1028  1994  [Refereed]

  • Oxygen-independent Photocleavage of Nucleic Acids with Macrocyclic Lanthanide Complexes

    Masami Kobayashi, Dai Yokotsuka, Naoya Takeda, Morio Yashiro, Makoto Komiyama

    Nucleic Acids, Symp. Ser.   31   159 - 160  1994  [Refereed]

  • Rare Earth Metal Ions for DNA Hydrolyses and Their Use to Artificial Nuclease

    Makoto Komiyama, Naoya Takeda, Tetsuro Shiiba, Yota Takahashi, Yoichi Matsumoto, Morio Yashiro

    Nucleosides and Nucleotides   13   1297 - 1309  1994  [Refereed]

  • Cerium(IV)-OligoDNA Hybrid as Highly Selective Artificial Nuclease

    Makoto Komiyama, Tetsuro Shiiba, Yota Takahashi, Naoya Takeda, Kazunari Matsumura, Teruyuki Kodama

    Supramol. Chem.   4   31 - 34  1994  [Refereed]

    DOI

  • Catalytically active species for CeCi3-induced DNA hydrolysis

    Makoto Komiyama, Teruyuki Kodama, Naoya Takeda, Jun Sumaoka, Tetsuro Shiiba, Yoichi Matsumoto, Morio Yashiro

    Journal of Biochemistry   115 ( 5 ) 809 - 810  1994  [Refereed]

     View Summary

    Cerium(III) chloride efficiently hydrolyzes DNA at pH 7 under aerobic conditions. A titration study showed that Ce(III) is oxidized to Ce(IV) in the reaction mixture and the resultant Ce(IV) is responsible for the DNA hydrolysis. © 1994 BY The Journal of Biochemistry.

    DOI PubMed

  • Cooperation of Lanthanum Ion and Non-Lanthanide Metal Ions for the Hydrolysis of Bis(4-Nitrophenyl)Phosphate

    Naoya Takeda, Makoto Irisawa, Makoto Komiyama

    J. Chem. Soc., Chem. Commun.   ( 24 ) 2773 - 2774  1994  [Refereed]

    Authorship:Lead author

     View Summary

    Lanthanum(III) ion and non-lanthanide metal ions [iron(III), manganese(III), tin(IV), indium(III) and gallium(III)] show a cooperative catalysis for the hydrolysis of bis(4-nitrophenyl)phosphate.

    DOI

  • DNA Hydrolysis by Cerium(IV) Does Not Involve either Molecular Oxygen or Hydrogen Peroxide

    Makoto Komiyama, Tetsuro Shiiba, Teruyuki Kodama, Naoya Takeda, Jun Sumaoka, Morio Yashiro

    Chem. Lett.     1025 - 1028  1994  [Refereed]

  • Oxygen-independent Photocleavage of Nucleic Acids with Macrocyclic Lanthanide Complexes

    Masami Kobayashi, Dai Yokotsuka, Naoya Takeda, Morio Yashiro, Makoto Komiyama

    Nucleic Acids, Symp. Ser.   31   159 - 160  1994  [Refereed]

  • Rare Earth Metal Ions for DNA Hydrolyses and Their Use to Artificial Nuclease

    Makoto Komiyama, Naoya Takeda, Tetsuro Shiiba, Yota Takahashi, Yoichi Matsumoto, Morio Yashiro

    Nucleosides and Nucleotides   13   1297 - 1309  1994  [Refereed]

  • Cerium(IV)-OligoDNA Hybrid as Highly Selective Artificial Nuclease

    Makoto Komiyama, Tetsuro Shiiba, Yota Takahashi, Naoya Takeda, Kazunari Matsumura, Teruyuki Kodama

    Supramol. Chem.   4   31 - 34  1994  [Refereed]

    DOI

  • Catalytically active species for CeCi3-induced DNA hydrolysis

    Makoto Komiyama, Teruyuki Kodama, Naoya Takeda, Jun Sumaoka, Tetsuro Shiiba, Yoichi Matsumoto, Morio Yashiro

    Journal of Biochemistry   115 ( 5 ) 809 - 810  1994  [Refereed]

     View Summary

    Cerium(III) chloride efficiently hydrolyzes DNA at pH 7 under aerobic conditions. A titration study showed that Ce(III) is oxidized to Ce(IV) in the reaction mixture and the resultant Ce(IV) is responsible for the DNA hydrolysis. © 1994 BY The Journal of Biochemistry.

    DOI PubMed

  • Lanthanide metal complexes for the hydrolysis of linear DNAs

    Tetsuro Shiiba, Koji Yonezawa, Naoya Takeda, Yoichi Matsumoto, Morio Yashiro, Makoto Komiyama

    Journal of Molecular Catalysis   84 ( 1 ) L21 - L25  1993.09  [Refereed]

     View Summary

    Lanthanum (III) and cerium (III) complexes of macrocyclic ligands efficiently hydrolyze the phosphodiester linkages in linear DNAs at pH 7.2 and 30-50°C. Both single-stranded and double-stranded DNAs are hydrolysed without specific base preference, yielding predominantly 3′-phosphate termini. The potential of these complexes as the catalytic centers of artificial hydrolytic nucleases are indicated. © 1993.

    DOI

  • Lanthanide metal complexes for the hydrolysis of linear DNAs

    Tetsuro Shiiba, Koji Yonezawa, Naoya Takeda, Yoichi Matsumoto, Morio Yashiro, Makoto Komiyama

    Journal of Molecular Catalysis   84 ( 1 ) L21 - L25  1993.09  [Refereed]

     View Summary

    Lanthanum (III) and cerium (III) complexes of macrocyclic ligands efficiently hydrolyze the phosphodiester linkages in linear DNAs at pH 7.2 and 30-50°C. Both single-stranded and double-stranded DNAs are hydrolysed without specific base preference, yielding predominantly 3′-phosphate termini. The potential of these complexes as the catalytic centers of artificial hydrolytic nucleases are indicated. © 1993.

    DOI

  • Molecular Design of Artificial Hydrolytic Nuclease and Ribonuclease

    Makoto Komiyama, Takuya Inokawa, Tetsuro Shiiba, Naoya Takeda, Koichi Yoshinari, Morio Yashiro

    Nucleic Acids, Symp. Ser.   29   197 - 198  1993  [Refereed]

  • Hydrolysis of oligodnas by lanthanide metal(III) chloride

    Makoto Komiyama, Yoichi Matsumoto, Nobuhiro Hayashi, Kazunari Matsumura, Naoya Takeda, Kimitsuna Watanabe

    Polymer Journal   25 ( 11 ) 1211 - 1214  1993  [Refereed]

    DOI

  • Site-Selective Hydrolysis of tRNA by Lanthanide Metal Complexes

    Nobuhiro Hayashi, Naoya Takeda, Tetsuro Shiiba, Morio Yashiro, Kimitsuna Watanabe, Makoto Komiyama

    Inorg. Chem.   32   5899 - 5900  1993  [Refereed]

    DOI

  • Molecular Design of Artificial Hydrolytic Nuclease and Ribonuclease

    Makoto Komiyama, Takuya Inokawa, Tetsuro Shiiba, Naoya Takeda, Koichi Yoshinari, Morio Yashiro

    Nucleic Acids, Symp. Ser.   29   197 - 198  1993  [Refereed]

  • Hydrolysis of oligodnas by lanthanide metal(III) chloride

    Makoto Komiyama, Yoichi Matsumoto, Nobuhiro Hayashi, Kazunari Matsumura, Naoya Takeda, Kimitsuna Watanabe

    Polymer Journal   25 ( 11 ) 1211 - 1214  1993  [Refereed]

    DOI

  • Site-Selective Hydrolysis of tRNA by Lanthanide Metal Complexes

    Nobuhiro Hayashi, Naoya Takeda, Tetsuro Shiiba, Morio Yashiro, Kimitsuna Watanabe, Makoto Komiyama

    Inorg. Chem.   32   5899 - 5900  1993  [Refereed]

    DOI

▼display all

Books and Other Publications

  • チップ基材の表面形状および性状が細胞に与える影響, 臓器チップの技術と開発動向

    今任 景一, 武田 直也( Part: Joint author)

    CMC  2018

Misc

  • Paper-based Double-layered Scaffold for Self-driven Perfusion Culture System

    Naoya Takeda

    Cellulose Communications   26 ( 4 ) 168 - 172  2019.12  [Invited]

  • 灌流培養のための紙を使った自律駆動型連続送液システムの設計

    有坂 慶紀, 尾﨑 愛, 今任 景一, 武田 直也

    バイオマテリアル –生体材料-   36 ( 2 ) 156 - 157  2018.04  [Refereed]

  • Thermoresponsive Polymer Grafted Micro-Structured Interfaces for Cell Separation

    Kenichi Nagase, Risa Shukuwa, Takahiro Onuma, Masayuki Yamato, Naoya Takeda, Teruo Okano

    Journal of Japanese Society for Biomaterials   36 ( 2 ) 158 - 159  2018.04  [Refereed]

  • 可逆的かつ長期安定的に高い細胞パターニング機能を実現するスピロピラン導入光応答性ソフト界面の創製

    有坂 慶紀, 何 迪, 今任 景一, 武田 直也

    Colloid & Interface Communication   43   20 - 23  2018  [Invited]

  • 配向化低密度「弦状」マイクロファイバー足場上で形成した高細胞密度筋管組織の機能構造解析

    田村健一, 峯口竜, 原雄介, 原口裕次, 清水達也, 岡野光夫, 武田直也

    再生医療   12  2013

    J-GLOBAL

  • エレクトロスピニング過程で化学架橋を施した配向化低密度「弦状」コラーゲンマイクロファイバー足場の作製と高細胞密度筋管組織の構築

    武田直也, 田村健一, 峯口竜, 原口祐次, 清水達也, 岡野光夫, 原雄介

    日本化学会講演予稿集   93rd ( 3 )  2013

    J-GLOBAL

  • 「弦状」に中空維持・配向化させたエレクトロスピン・コラーゲンファイバー足場での三次元筋管形成誘導

    田村健一, 峯口竜, 原雄介, 原口裕次, 清水達也, 岡野光夫, 武田直也

    再生医療   11  2012

    J-GLOBAL

  • Navigation of Neurite Elongation and Fabrication of the Networks of Neural Cells with Fine Resolution on the Novel Cell Culture Surface Patterned by Electron Beam Lithography

    N. Takeda, N. Yoshino, Y. Edagawa, N. Shimamoto

    Neurosci. Res.   65 ( Suppl. 1 ) e219  2010

  • Regulation of the Outgrowth Position of Neurites in Cell Body and the Direction of Their Elongation for PC12 Cells in Three Dimensional Hydrogel Culture System with Feeble Flow of the Solvent

    N. Takeda, J. Uehara, Y. Edagawa

    Neurosci. Res.   64 ( Suppl. 1 ) S132  2009

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Industrial Property Rights

  • 光に応答して可逆的に表面相分離構造を変える細胞培養用基材

    武田 直也, 今任 景一, 坂野 誠人, 松下 武司, 福士 英明, 謝 小毛

    Patent

  • チキソトロピー性を有するゲルを用いる多層3次元細胞培養足場システム

    武田 直也, 今任 景一, 米田 憲司, 田中 智, 神野 和人, 後居 洋介

    Patent

  • 導電性ポリマー層と細胞接着用ポリマー層とを含む多層膜からなる細胞培養用基材

    武田 直也, 今任 景一, 田中 啓太, 中谷 美沙, 謝 小毛, 福士 英明

    Patent

  • 細胞捕捉装置及びそれを利用した細胞操作方法

    荒川 貴博, 武田 直也, 山口 佳則, 枝川 義邦

    Patent

  • タンパク質又は遺伝子導入用試薬

    5403324

    武岡 真司, 武田 直也, 胡桃坂 仁志, 坂根 勲, 池ヶ谷 菜海子, 小幡 洋輔, 齋藤 俊介

    Patent

Other

  • [1] (財)カシ...

     View Summary

    [1] (財)カシオ科学振興財団研究協賛事業,「神経筋接合部により運動機能を制御するin vitro再生骨格筋組織の構築」,2011/11/1〜2013/10/31.
    [2] (財)松籟科学技術振興財団・2009年度研究助成,「細胞の分化・極性を制御するナノ構造化高分子培養基板システムの構築」,2010/4/1〜2011/3/31.
    [3] (財)熊谷科学技術振興財団・平成16年度試験研究助成,「温度応答性高分子を基盤としたインテリジェントな遺伝子デリバリーシステムの構築とその応用」,2004/9/1〜2006/3/31.

Awards

  • Encouragement Award at 15th Annual Meeting of Nano biomedical Society

    2020.11   Nano Biomedical Society  

  • Poster Presentation Award 2019 for Excellent Research, 9th CSJ Chemistry Festa 2019

    2019.10   The Chemical Society of Japan  

    Winner: Masato SAKANO, Haruka NISHIDA, idually awarded

  • Highlight Paper of the 41st Annual Meeting of the Japanese Society for Biomaterials

    2019   Japanese Society for Biomaterials  

    Winner: Mari Nagasawa, Yuji Tsuchido, Naoya Takeda

  • Excellent Presentation Award, 2nd G'L'owing Polymer Symposium in KANTO

    2019   Kanto Branch, The Society of Polymer Science, Japan  

    Winner: Yutaro Miyake

  • Excellent Poster Presentation Award at 68th Symposium on Macromolecules

    2019   The Society of Polymer Science, Japan  

    Winner: Haruka Nishida

  • Reviewer Certificate

    2018   Polymer International  

    Winner: Naoya Takeda

  • Teaching Award, 2018 Autumn

    2018   Waseda University  

    Winner: Naoya Takeda

  • Poster Award

    2017   The 2nd International Symposium on Bio-Therapeutics Delivery  

    Winner: Risa Shukuwa

  • Excellent Poster Presentation Award at 66th Symposium on Macromolecules

    2017   The Society of Polymer Science, Japan  

    Winner: Hidekazu Nakajima

  • CSJ Poster Presentation Award 2017 for Excellent Research

    2017   7th CSJ Chemistry Festa 2017  

    Winner: Masao Maruyama

  • CSJ Poster Presentation Award 2017 for Excellent Research

    2017   7th CSJ Chemistry Festa 2017  

    Winner: Kenji Yoneda

  • Bio4Apps 2017 Best Award

    2017   International Conference on BioSensors, BioElectronics, BioMedical Devices, BioMEMS/NEMS & Applications (Bio4Apps 2017)  

    Winner: Masao Maruyama

  • Teaching Award, Waseda University

    2017   Waseda University   Organic Chemistry A

    Winner: Naoya Takeda

  • Publicity Award, Publicity Committee, The Society of Polymer Science, Japan

    2017   Publicity Committee, The Society of Polymer Science, Japan   チキソトロピー性セルロースゲルを培養場とした多層ファイバー中での筋管形成

    Winner: Keiichi Imato, Kenji Yoneda, Tomo Tanaka, Naoya Takeda

  • Highlight Paper of the 39th Annual Meeting of the Japanese Society for Biomaterials

    2017   Japanese Society for Biomaterials  

    Winner: Kenichi Nagase, Lisa Shukuwa, Takahiro Onuma, Masayuki Yamato, Naoya Takeda, Teruo Okano

  • Excellent Poster Presentation Award at 65th Symposium on Macromolecules

    2016   The Society of Polymer Science, Japan  

    Winner: Risa Shukuwa

  • Excellent Poster Presentation Award at 65th Symposium on Macromolecules

    2016   The Society of Polymer Science, Japan  

    Winner: Keita Tanaka

  • Poster Presentation Award for the Excellent Research

    2016   6th CSJ Chemistry Festa 2016   6th CSJ Chemistry Festa 2016

    Winner: Kazuho Nagata

  • Poster Presentation Award for the Excellent Research

    2015   5th CSJ Chemistry Festa 2015  

    Winner: Miki Sawatari

  • Poster Presentation Award for the Excellent Research

    2015   5th CSJ Chemistry Festa 2015  

    Winner: Di He

  • The Best Poster Presentation Award for the Most Excellent Research

    2015   5th CSJ Chemistry Festa 2015  

    Winner: Kenichi Masuda

  • Excellent Poster Presentation Award

    2015   The International Symposium on Nanoarchitectonics for Mechanobiology  

    Winner: Miki Sawatari

  • Highlight Paper of the 37th Annual Meeting of the Japanese Society for Biomaterials

    2015   Japanese Society for Biomaterials  

    Winner: Ai Ozaki, Yoshinori Arisaka, Naoya Takeda

  • Excellent Presentation Award for the Student at the 25th Inteligent Materials/Systems Symposium

    2015   The Society of Non-Traditional Technology, Intelligent Materials/Systems Forum  

    Winner: Ai Ozaki

  • Poster Presentation Award for the Excellent Research

    2014   4th CSJ Chemistry Festa 2014  

    Winner: Tetsuya Kurebayashi

  • Teaching Award, Waseda University

    2014   Waseda University   Analytical Chemistrry A

    Winner: Naoya Takeda

  • Bio4Apps 2013 Best Award

    2013   International Conference on BioSensors, BioElectronics, BioMedical Devices, BioMEMS/NEMS and Applications 2013 & 5th Sensing Biology Symposium  

    Winner: Hitomi Oku

  • Poster Presentation Award for the Excellent Research

    2013   3rd CSJ Chemistry Festa 2013  

    Winner: Yutaro Nakamura

  • Poster Presentation Award for the Excellent Research

    2012   2nd CSJ Chemistry Festa 2012  

    Winner: Aya Tsubokura

  • ICBS2013 Biomaterials Science Poster Prize (sponsored by RSC Publishing)

    2012   2nd International Conference on Biomaterials Science (ICBS2013)  

    Winner: Aya Tsubokura

  • Biotechnology and Bioengineering, Top 50 Reviewers of 2012

    2012  

    Winner: Naoya Takeda

  • Top 50 Reviewer

    2012   Biotechnology & Bioengineering  

    Winner: Naoya Takeda

  • Poster Presentation Award for the Excellent Research

    2011   1st CSJ Chemistry Festa 2011  

    Winner: Kenichi Tamura

  • Poster Presentation Award for the Excellent Research

    2011   1st CSJ Chemistry Festa 2011  

    Winner: Yoshifumi Kawagishi

  • Excellent Presentation Award for the Student

    2011   The Society of Non-Traditional Technology, Intelligent Materials/Systems Forum  

    Winner: Kenichi Tamura

  • Grant Award, 27th Shorai Foundation for Science and Technology

    2009   Shorai Foundation for Science and Technology   細胞の分化・極性を制御するナノ構造化高分子培養基板システムの構築

    Winner: Naoya Takeda

▼display all

Research Projects

  • チキソトロピー性セルロースナノファイバーゲルを基盤とした生体組織作製系の創成

    Project Year :

    2018.06
    -
    2020.03
     

  • チキソトロピー性セルロースナノファイバーゲルを基盤とした生体組織作製系の創成

    科学研究費助成事業(日本学術振興会), 早稲田大学  科学研究費助成事業(学術研究助成基金助成金)

    Project Year :

    2018.06
    -
    2020.03
     

     View Summary

    独自開発したマイクロ流体デバイスを用いて、生体適合性・チキソトロピー性・高いアスペクト比のセルロースナノファイバー(CNF)ゲルを細胞培養層とした、多層ゲルファイバー培養場を作製し、包埋した細胞を分化・成熟させることで長大で複雑なヒト筋組織・血管組織を構築する

  • 組織構築を誘導するセルロースナノファイバー培養場の開発

    科学技術振興機構  研究成果展開事業 研究成果最適展開支援プログラム(A-STEP)

    Project Year :

    2018.09
    -
    2019.08
     

     View Summary

    様々な特性のCNFを用いて細胞からの血管や筋組織形成への影響を解明し、最適な培養場の作製方法を確立する。

  • Development of polymeric materials with a highly sensitive mechanoprobe based on charge-transfer interaction and application to mechanobiology

    Project Year :

    2016.08
    -
    2018.03
     

     View Summary

    In this study, we tried to develop a highly sensitive mechanoprobe based on charge transfer (CT) interaction between fluorescent molecules that reversibly dissociate and associate, and incorporate it into polymer chains, in order to reversibly detect microstress in polymeric materials. The designed probe and polymeric materials with it were successfully synthesized, but the reversible detection of microstress could not be achieved. On the other hand, when CT interaction was utilized as reversible physical cross-linking points in polymeric materials (elastomers), the elastomers showed self-healing in mild environments and could be used as cell culture scaffolds

  • 三次元層流マイクロ流体デバイスによる同軸多層ゲルファイバーを足場とした成熟血管組織作製システムの創成

    公益財団法人 カシオ科学振興財団  第33回研究助成

    Project Year :

    2015.12
    -
    2016.11
     

     View Summary

    新型の高性能な同軸多層型の三次元層流PDMSマイクロ流体デバイスを用いて、 タンパク質またはペプチドを組み込み細胞接着性を増大させたゲルファイバー材料を開発する。これを細胞培養場に用いて、三次元の生体組織を作製する。

  • 光刺激により物性とナノ微細構造を変化させる非分解放出型機能性高分子表面の創成と細胞マニピュレーションへの応用

    公益財団法人 泉科学技術振興財団  平成27年度研究助成

    Project Year :

    2015.10
    -
    2016.09
     

     View Summary

    スピロピランを導入した高分子材料を用いて、光刺激で材料の表面特性を変化できる新たな細胞培養器材の開発を行う。この光応答性培養器材により、まず細胞接着と剥離の制御を行う。

  • Advanced Micro Fluidic Engineering and Its Applications for High Sensitive Quantitative Measurements of Biomolecules

    Project Year :

    2011.05
    -
    2016.03
     

     View Summary

    In this study, the noble microfluidic device technology was developed in order to enable the optical high sensitive quantitative measurement of the micro luminescent sample as follows. 1) The micro droplet manufacturing technology of the various size. 2) The control technology of the micro flow. 3) The passive sorting technology of the micro droplet. Next, the measurement of the micro luminescent samples by systematizing microfluidic device was realized. 1) Insert a living cell to a droplet one by one, and analyze the gene of the environmental microbe. 2) Observation cellular growth and evaluate enzymatic reaction activity. Optical quantitative measurement was realized by this study

  • 神経筋接合部により運動機能を制御するin vitro再生骨格筋組織の構築

    公益財団法人 カシオ科学振興財団  第2回研究協賛金

    Project Year :

    2012.11
    -
    2013.10
     

     View Summary

    再生医療や生理・病理学解析への応用に資する神経系・運動器系の再生複合組織の構築に向けて、「神経束 → 筋 → 腱骨移行部」の各構成組織について、細胞から三次元の再生組織をin vitroで構築するための基盤工学技術を開発した。

  • 基質界面の微細構造による細胞遊走の整流化制御と形質転換因子としての評価

    Project Year :

    2011.04
    -
    2013.03
     

     View Summary

    細胞が基質へ接触や接着をしさらに伸展や遊走など運動をする際、接着や細胞骨格に関与するタンパク質群は時空間的に変動し、接着斑や細胞骨格を介したメカノストレス伝達に影響すると考えられる。本研究では、電子線リソグラフィー技術を基盤とした独自の細胞パターニングシステムにより作製した微細パターン構造のみでヒト骨髄由来間葉系幹細胞(hbmMSC)の接着能を制御し、遊走や運動ひいては増殖や分化の各挙動の制御を目指した。これら挙動変化と接着斑やアクチンストレスファイバー形成との相関も評価した。連続的にV字型マイクロパターンを設けた一方向細胞遊走誘導システムでは(遊走速度0.2um/min)、メカノセンシングに関与するとされるアクチンキャップ形成を見出した。5日間培養時の定量qRT-PCRによる分化挙動評価では、平面培養との比較で、ALCAM、CD44、CD29の各幹細胞マーカーmRNAの発現低下傾向や神経分化マーカーNestinの発現亢進が見出され、一方向遊走運動と

  • Development of the Novel Nano Trench-Patterned Culture Surface for Initiating Differentiation of Mesenchymal Stem Cell

    Project Year :

    2010.04
    -
    2013.03
     

     View Summary

    The novel cell patterning system was developed by utilizing the electron beam (EB) lithography and the EB-reactive polymer resists. And as a new method, we achieved inducing differentiation of mesechymal stem cell (MSC) just by culturing at the nano-topographic pattern of a single linear groove in 500 nm width, without transfecting any DNA and/or applying any biosignaling factor like concentional biological methods. MSCs were extraordinarily streched along the groove, presumably accompanying mechanical stress, and markedly increased gene expression of the neural markers and the osteogenic markers

  • An in vivo gene expressing system induced by external thermal stimuli

     View Summary

    A thermo-responsive polymer was synthesized from N-isopropyl acrylamide, 2-(dimethylamino)ethyl methacrylate, and butylmethacrylate. This copolymer possessed a phase transition temperature at 25℃. Behaviors of its complexation with plasmid DNA were analyzed. When this polymer was complxed with DNA at a N/P charge ratio of 4, zeta-potencial of the formed complex was +14.1mV. Binding affinity of this polymer to DNA was revealed to change on temperature stimuli ; lower affinity at higher temperature. This induced change of affinity could contribute to DNA release from the complex on temperature drop. This affinity change for this polymer was larger than that obtained in previously studied polymers that synthesized with lower contents of 2-(dimethylamino)ethyl methacrylate and butylmethacrylate than the polymer of the present study.In vitro gene expression assay revealed that this thermo-responsive polymer exhibited thermo-responsive gene expression ; two-fold gene expression upon cooling at 20℃ for 3h. This polymer showed much higher level of gene expression than the previously studied polymer. Furthermore, cytotoxicity of this polymer complex was considerably low such as 80% bioa

  • 温度応答性高分子キャリアーの遺伝子デリバリーシステムへの応用とその制御機構の解明

     View Summary

    温度に応答して相転移を起こすN-isopropylacrylamide(IPAAm)とカチオン性の3級アミノ基を有する2-(dimethylamino)ethyl methacrylate(DMAEMA)、さらに疎水的な分子であるbutylmethacrylate(BMA)の3つのユニットからなる直鎖状のrandom copolymer(IP-20D-10B、合成時のDMAEMAおよびBMAのモル比がそれぞれ20,10%)について、COS-1細胞に対する遺伝子導入効率を動力学的に解析した。遺伝子にはβ-galactosidaseをコードしたplasmid DNAを用い、これを96well plateの1wellに対し0.1μg加えた。既往研究と比較して、この濃度は2分の1から10分の1という低い条件である。この低dose条件でも、IP-20D-10BはCOS-1細胞に効率的に遺伝子を導入することができる。IP-20D-10Bとplasmid DNAの比を変化させて遺伝子導入効率と細胞毒性へ与える影響を検討した。高分子キャリアーの比率を増加させるに従い遺伝子導入効率は増大し、N/P比(高分子のアミノ基とDNAのリン酸基のモル比)10/1で飽和に達した。しかし、細胞毒性も徐々に増大し、N/P=10では10%と顕著になった。一方

  • 温度応答性培養皿への遺伝子固定化と熱刺激による界面からのデリバリー制御

     View Summary

    温度に応答して相転移を起こすN-isopropylacrylamide(IPAAm)を主として、カチオン性と疎水性の3つのユニットからなる直鎖状のランダム共重合体(IP-20D-10B)について、293細胞に対する遺伝子導入効率を評価した。導入する遺伝子には、GFPをコードしたプラスミドDNAを用いた。まず定法の培地に添加する方法でIP-20D-10B/DNA複合体を293細胞に加えた所、confluentで細胞分裂が抑制された状態であっても40%程度(t=4d)の高い遺伝子導入効率が得られた。続いて、IPAAmにカルボキシル基を導入した誘導体(CIPAAm)を合成し、IPAAmとCIPAAmを培養基材表面に電子線でグラフトして、温度応答性培養皿を作製した(CIPAAmのモル分率は1,3,5%)。ここに、正電荷過剰のIP-20D-10B/DNA複合体を静電的に固定し、さらに293細胞を播種して細胞の基底面からの遺伝子導入を試みた。CIPAAmのモル比の増大に伴って細胞接着性が低下し、5%CIPAAm培養皿では培養1日目で細胞の凝集が観測された。遺伝子の導入効率は、0、01%以下ときわめて低かった。

  • 細胞の分化・極性を制御するナノ構造化高分子培養基板システムの構築

    財団法人 松籟科学技術振興財団  第27回松籟科学技術振興財団研究助成

▼display all

Presentations

  • Mechano-stress Sensing by Fluorescent Visualization of Supramolecular Polymers Possessing Charge-transfer Complex

    Yuji Tsuchido, Ryota Yamanaka, Hidekazu Nakajima, Naruki Ito, Kenta Iitani, Keiichi Imato, Naoya Takeda

    RSC Tokyo International Conference 2020 

    Event date:
    2020.12
     
     
  • Photocontrol of Wettability and Cell Adhesion on Nanofilm Surfaces of Copolymers Having Spiropyran Molecules and PEG Block

    Masafumi Mukasa, Mitsuya Yamamoto, Kenta Iitani, Yuji Tsuchido, Naoya Takeda

    3rd Glowing Polymer Symposium in KANTO 

    Presentation date: 2020.11

    Event date:
    2020.11
     
     
  • バイオマス由来ナノ材料三次元培養場を用いた管腔構造をもつ血管組織の作製

    角田敬正, 西田春霞, 土戸優志, 関口哲志, 庄子習一, 武田直也

    第15回ナノバイオメディカル学会大会 

    Presentation date: 2020.11

    Event date:
    2020.11
     
     
  • 紙とマイクロファイバーの二層構造基材を用いた気液界面培養による腸管上皮組織の構築

    大貫真依, 長澤真理, 土戸優志, 武田直也

    第15回ナノバイオメディカル学会大会 

    Presentation date: 2020.11

    Event date:
    2020.11
     
     
  • パターン化温度応答性培養基材を用いた筋衛星細胞からの配向化ヒト骨格筋組織の作製

    石山果穂, 高橋宏信, 清水達也, 武田直也

    第10回CSJ化学フェスタ2020 ―日本化学会秋季事業― 

    Presentation date: 2020.10

    Event date:
    2020.10
     
     
  • 電荷移動錯体を主鎖に導入した微小応力を蛍光検出可能なエラストマーの創製

    伊藤成輝, 山中凌大, 飯谷健太, 土戸優志, 今任景一, 武田直也

    第10回CSJ化学フェスタ2020 ―日本化学会秋季事業― 

    Presentation date: 2020.10

    Event date:
    2020.10
     
     
  • 直列に結合した電子ドナー・アクセプターの電荷移動錯体を主鎖にもつメカノストレス・センシング高分子材料の機能

    土戸優志, 山中凌大, 伊藤成輝, 中島英和, 飯谷健太, 今任景一, 武田直也

    日本分析化学会第69年会 

    Presentation date: 2020.09

    Event date:
    2020.09
     
     
  • スピロピランの光異性化を駆動力として構造・物性を動的転換する表面設計と培養基材応用

    武田直也, 渡邉里奈, 三宅裕太郎, 坂野誠人, 梅野春花, 今任景一, 土戸優志

    第69回高分子討論会 

    Presentation date: 2020.09

    Event date:
    2020.09
     
     
  • 電荷移動錯体を利用した高分子材料への力学的負荷検出

    土戸優志, 山中凌大, 今任景一, 武田直也

    第80回分析化学討論会 

    Presentation date: 2020.05

    Event date:
    2020.05
     
     
  • Real-time imaging of transcutaneous ethanol using biofluorometric gas-imaging system (Sniff-cam) with body shape fitting interface for 2D enzyme immobilized mesh sheet

    Kenta Iitani, Naoki Mizukoshi, Koji Toma, Takahiro Arakawa, Naoya Takeda, Kohji Mitsubayashi

    18th International Meeting on Chemical Sensors 2020 

    Presentation date: 2020.05

  • ヒトiPS細胞由来運動神経と筋の複合組織における神経筋接合部の評価

    Fumiko Oikawa, Hironobu Takahashi, Tatsuya Shimizu, Naoya Takeda

    Presentation date: 2020.03

  • 同軸二層構造セルロースナノファイバーゲルでの三次元組織の構築と移植応用に向けた動態解析

    Naoya Takeda, Haruka Nishida, Takamasa Tsunoda, Hidekazu Sekine, Doon-Hyun Yun, Tetsushi Sekiguchi, Shuichi Shoji, Tatsuya Shimizu

    第19回日本再生医療学会総会 

    Presentation date: 2020.03

  • Fabrication of Engineered 3D Tissues in Biomimetic Environments Using Microfiber-based Scaffolds

    Naoya TAKEDA, Mari Nagasawa, Mai Onuki, Yuji Tsuchido  [Invited]

    International Conference on BioSensors, BioElectronics, BioMedical Devices, BioMEMS/NEMS & Applications (Bio4Apps 2019) 

    Presentation date: 2019.12

  • フィブリンゲルを用いて作製した 骨格筋組織 の 筋 損傷 モデルとしての応用を目指した基礎検討

    Hironobu Takahashi, Hazuki Tanaka, Kaho Ishiyama, Tatsuya Shimizu, Naoya Takeda

    Presentation date: 2019.11

  • 光駆動型動的ソフト界面における可逆的相分離微細構造変化の機構と細胞応答挙動の解析

    Masato Sakano, Keiichi Imato, Yuji Tsuchido, Naoya Takeda

    Presentation date: 2019.11

  • 二層基材上での気液界面培養による粘液層と三次元構造を有した腸管上皮モデル組織の構築

    Mari Nagasawa, Yuji Tsuchido, Naoya Takeda

    Presentation date: 2019.11

  • Dynamic Photoconversion of Nanoscale Phase separated Structures Formed on the Spiropyran introduced Polymer Thin Film

    Yutaro Miyake, Masato Sakano, Keiichi Imato, Naoya Takeda

    2nd GLowing Polymer Symposium in KANTO 

    Presentation date: 2019.11

  • Design and Preparation of Polymer Materials with Stress Sensing Function by Charge Transfer Complex

    Ryota YAMANAKA, Hidekazu NAKAJIMA, Yuji TSUCHIDO, Keiichi IMATO, Naoya TAKEDA

    2nd GLowing Polymer Symposium in KANTO 

    Presentation date: 2019.11

  • Photomanipulation of dynamic behaviors of polymer materials and cells at nanobio interfaces

    Naoya Takeda  [Invited]

    Presentation date: 2019.11

  • 可逆的な表面相分離微細構造の光転換と細胞挙動制御の動的解析

    Msato Sakano, Keiichi Imato, Yuji Tsuchido, Naoya Takeda

    9th CSJ Chemistry Festa 2019 

    Presentation date: 2019.10

  • ナノスケールの相分離構造形成と光転換制御を実現する高分子薄膜材料の創製

    Yutaro Miyake, Masato Sakano, Keiichi Imato, Yuji Tsuchido, Naoya Takeda

    9th CSJ Chemistry Festa 2019 

    Presentation date: 2019.10

  • 光刺激に応答して相分離構造と細胞接着挙動を転換するナノ薄膜材料の表面解析

    Haruka Umeno, Masato Sakano, Keiichi Imato, Yuji Tsuchido, Naoya Takeda

    9th CSJ Chemistry Festa 2019 

    Presentation date: 2019.10

  • 分子内電荷移動錯体で応力を蛍光検出する高分子材料の創製

    Ryota Yamanaka, Hidekazu Nakajima, Yuji Tsuchido, Keiichi Imato, Naoya Takeda

    9th CSJ Chemistry Festa 2019 

    Presentation date: 2019.10

  • 三次元培養足場としてのCNFゲルの材料物性評価と再生組織構築

    Haruka Nishida, Tomo Tanaka, Doon-Hyun Yun, Yuji Tsuchido, Keiichi Imato, Tetsushi Sekiguchi, Shuichi Shoji, Naoya Takeda

    9th CSJ Chemistry Festa 2019 

    Presentation date: 2019.10

  • 効率的筋管形成を可能とする導電性コアシースファイバー足場の構造機能解析

    Misa Nakaya, Yuji Tsuchido, Naoya Takeda

    9th CSJ Chemistry Festa 2019 

    Presentation date: 2019.10

  • 紙とマイクロファイバーからなる新規培養基材での気液界面培養による三次元腸モデル組織の作製

    Mari Nagasawa, Yuji Tsuchido, Naoya Takeda

    9th CSJ Chemistry Festa 2019 

    Presentation date: 2019.10

  • Correlation study on structures, physical properties, and functions of cellulose nanofiber gels as three-dimensional culture scaffolds in engineered tissues fabrication

    Haruka NISHIDA, Tomo TANAKA, Dong Hyun YOON, Yuji TSUCHIDO, Keiichi IMATO, Tetsushi SEKIGUCHI, Shuichi SHOJI, Naoya TAKEDA

    Presentation date: 2019.09

  • Correlation Study on Structures, Physical Properties, and Functions of Polymer Materials Containing a Charge Transfer Complex as Mechanoprobe in the Main Chain

    Ryota Yamanaka, Hidekazu Nakajima, Yuji Tsuchido, Keiichi Imato, Naoya Takeda

    Presentation date: 2019.09

  • Fabrication of engineered tissues using the functional two-layer culture substrates consisting of paper and electrospun polymer microfibers

    Naoya TAKEDA, Mari NAGASAWA, Yuji Tsuchido

    Presentation date: 2019.09

  • Dynamic molecular motion-induced cellular responses in microstructure changes of photoresponsive polymer surface

    Naoya TAKEDA, Masato SAKANO, Yutaro MIYAKE, Haruka UMENO, Yuji TSUCHIDO, Keiichi IMATO  [Invited]

    Presentation date: 2019.09

  • Fabrication of engineered tissues using multifunctional bilayer culture scaffold composed of electrospun polymer microfibers and paper

    Naoya Takeda, Mari Nagasawa, Masao Maruyama

    Presentation date: 2019.07

  • TEMPO-oxidized cellulose nanofiber gel scaffold for fabricating three-dimensional engineered tissues

    Naoya Takeda, Haruka Nishida, Tomo Tanaka, Dong Hyun Yoon, Keiichi Imato, Tetsushi Sekiguchi, Shuichi Shoji

    Presentation date: 2019.07

  • Perfect photoswitching of phase-separated structures formed by block copolymer thin films and spatio-temporal control of cell behavior

    Keiichi Imato, Masato Sakano, Naoya Takeda

    Presentation date: 2019.05

  • Control of nanophase-separated structures formed by photoresponsive block copolymer thin films

    Yutaro MIYAKE, Masato SAKANO, Keiichi IMATO, Naoya TAKEDA

    Presentation date: 2019.05

  • Preparation of polymer material containing charge transfer complex of fluorescent molecules in the main chain for stress sensing

    Ryota YAMANAKA, Hidekazu NAKAJIMA, Keiichi IMATO, Naoya TAKEDA

    Presentation date: 2019.05

  • 紙を基材に用いた気液培養系による腸管上皮組織の作製

    武田直也, 長澤真理, 今任景一

    第18回日本再生医療学会総会 

    Presentation date: 2019.03

  • 分子構造を可逆的に光転換できる動的な培養表面からの単一細胞の剥離回収

    武田直也, 山本光哉, 今任景一

    第18回日本再生医療学会総会 

    Presentation date: 2019.03

  • Fabrication of Engineered Tissues by Cell Culture in Cellulose Nanofiber Hydrogel

    Naoya Takeda  [Invited]

    日本化学会 第99回春季年会(2019) 

    Presentation date: 2019.03

  • Reversible and non-degradable photocontrol of oppositely changing wettability and cell patterning of the blockcopolymer surface driven by spiropyran isomerization

    Naoya Takeda

    1st GLowing Polymer Symposium in KANTO 

    Presentation date: 2018.12

  • 光刺激でLCSTが変化するスピロピラン導入PNIPAmブラシ表面の創製と細胞挙動制御

    渡邉里奈, 長田和歩, 今任景一, 武田直也

    第40回日本バイオマテリアル学会大会 

    Presentation date: 2018.11

  • 紙とマイクロファイバーの二層基材を用いた気液界面培養による腸管上皮モデル組織の作製

    長澤真理, 今任景一, 武田直也

    第40回日本バイオマテリアル学会大会 

    Presentation date: 2018.11

  • 光応答性高分子足場を用いた可視光刺激による単一細胞の剥離制御

    山本 光哉, 今任 景一, 武田 直也

    第40回日本バイオマテリアル学会大会 

    Presentation date: 2018.11

  • 神経筋組織の作製を目的とした温度応答性パターン化基材による配向制御とフィブリンゲルを用いた培養手法

    及川 富美子, 高橋 宏信, 清水 達也, 武田 直也

    第40回日本バイオマテリアル学会大会 

    Presentation date: 2018.11

  • 光刺激によるLCST制御が可能なPNIPAmポリマーブラシ表面の創製と細胞接脱着操作

    渡邉里奈, 長田和歩, 今任景一, 武田直也

    第8回CSJ化学フェスタ2018 ―日本化学会秋季事業― 

    Presentation date: 2018.10

  • 紙と高分子マイクロファイバーの二層基材を用いた気液界面培養による腸管上皮モデル組織の作製

    長澤真理, 今任景一, 武田直也

    第8回CSJ化学フェスタ2018 ―日本化学会秋季事業― 

    Presentation date: 2018.10

  • 微小力検知を志向した分子内電荷移動錯体を基盤とするメカノプローブの設計

    中島英和, 山中凌大, 今任景一, 武田直也

    第8回CSJ化学フェスタ2018 ―日本化学会秋季事業― 

    Presentation date: 2018.10

  • スピロピランとPEGの相互作用変化に基づく光応答性表面の細胞接脱着制御

    山本光哉, 今任景一, 武田直也

    第8回CSJ化学フェスタ2018 ―日本化学会秋季事業― 

    Presentation date: 2018.10

  • UV/可視光刺激で可逆的かつ完全に相分離構造が消失/再形成するジブロック共重合体薄膜の創製と動的細胞挙動制御

    坂野誠人, 今任景一, 武田直也

    第8回CSJ化学フェスタ2018 ―日本化学会秋季事業― 

    Presentation date: 2018.10

  • 温度応答性高分子パターン化基材での共培養による骨格筋と運動神経の配向化複合組織の作製

    及川富美子, 高橋宏信, 清水達也, 武田直也

    第8回CSJ化学フェスタ2018 ―日本化学会秋季事業― 

    Presentation date: 2018.10

  • Effective development of neuromuscular junction in coculture with hiPS cell-derived motor neurons and oriented engineered muscle tissue

    Fumiko Oikawa, Hironobu Takahashi, Tatsuya Shimizu, Naoya Takeda

    5th Tissue Engineering and Regenerative Medicine international Society World Congress  (Kyoto) 

    Presentation date: 2018.09

  • Dual stimuli-responsive PIPAAm-PDMS surfaces for controlling cell attachment and detachment

    Yoshikatsu Akiyama, Miki Matsuyama, Naoya Takeda, Masayuki Yamato, Teruo Okano

    5th Tissue Engineering and Regenerative Medicine international Society World Congress  (Kyoto) 

    Presentation date: 2018.09

  • Enhanced myotube formation on the oriented coaxial coresheath microfiber scaffold embedding high conductive polymer without applied voltage

    Misa Nakaya, Keita Tanaka, Keiichi Imato, Naoya Takeda

    5th Tissue Engineering and Regenerative Medicine international Society World Congress  (Kyoto) 

    Presentation date: 2018.09

  • Perfusion culture system using paper-based bilayer scaffold with porous microfibers for effective myotube formation

    Masao Maruyama, Keiichi Imato, Naoya Takeda

    5th Tissue Engineering and Regenerative Medicine international Society World Congress  (Kyoto) 

    Presentation date: 2018.09

  • Fabrication of long myotube bundles in three-dimensional gel scaffolds with oriented cellulose nanofibers

    Keiichi Imato, Kenji Yoneda, Tomo Tanaka, Dong, Hyun Yoon, Tetsushi Sekiguchi, Shuichi Shoji, Naoya Takeda

    5th Tissue Engineering and Regenerative Medicine international Society World Congress  (Kyoto) 

    Presentation date: 2018.09

  • Fabrication of long luminal tissues in three-dimensional gel scaffolds with oriented cellulose nanofibers

    Tomo Tanaka, Keiichi Imato, Kenji Yoneda, Dong Hyun Yoon, Tetsushi Sekiguchi, Shuichi Shoji, Naoya Takeda

    5th Tissue Engineering and Regenerative Medicine international Society World Congress  (Kyoto) 

    Presentation date: 2018.09

  • スピロピラン導入ブロックとPEGからなる共重合体の光応答性細胞操作の機構解析

    山本光哉, 今任景一, 武田直也

    第67回高分子討論会 

    Presentation date: 2018.09

  • 分子内電荷移動錯体を有する高分子蛍光プローブの設計

    中島英和, 山中凌大, 今任景一, 武田直也

    第67回高分子討論会 

    Presentation date: 2018.09

  • スピロピラン導入高分子の光誘起相転移で動的に機能するバイオインターフェースの開発

    武田直也, 山本光哉, 坂野誠人, 三宅裕太郎, 梅野春花, 今任景一  [Invited]

    第67回高分子討論会 

    Presentation date: 2018.09

  • 三次元培養場の配向ナノ繊維が誘導する長大な再生組織構築

    今任景一, 米田憲司, 田中 智, 武田直也

    第67回高分子学会年次大会 

    Presentation date: 2018.05

  • スピロピラン導入高分子表面におけるミクロ相分離構造の 完全光スイッチングと細胞培養基材応用

    坂野誠人, 今任景一, 武田直也

    第67回高分子学会年次大会 

    Presentation date: 2018.05

  • Paper-based Perfusion Culture System Combined with Microfiber Technology

    Naoya Takeda  [Invited]

    IEEE The International Conference on Nano/Micro Engineered and Molecular Systems 2018  (Singapore) 

    Presentation date: 2018.04

  • 電荷移動錯体を架橋点に用いた自己修復性エラストマーの培養基材応用

    中島 英和, 今任 景一, 武田 直也

    第27回インテリジェント材料・システムシンポジウム 

    Presentation date: 2018.01

  • ソフト界面の精密設計が実現する細胞接脱着の光スイッチング制御

    山本 光哉, 今任 景一, 武田 直也

    第27回インテリジェント材料・システムシンポジウム 

    Presentation date: 2018.01

  • マイクロ・ナノ加工化高分子足場による細胞からの組織作製

    武田直也  [Invited]

    第16回高分子ナノテクノロジー研究会講座  高分子学会ナノテクノロジー研究会

    Presentation date: 2018.01

  • Effective myotube formation induced by shear stress in perfusion culture using porous microfiber scaffold

    Masao Maruyama, Naoya Takeda

    International Conference on BioSensors, BioElectronics, BioMedical Devices, BioMEMS/NEMS & Applications (Bio4Apps 2017) 

    Presentation date: 2017.12

  • Fabrication of Muscle Tissue in Thixotropic Cellulose Nanofiber Gel Scaffold Using MEMS Technology

    Naoya Takeda, Kenji Yoneda, Tomo Tanaka, Dong Hyun Yoon, Tetsushi Sekiguchi, Shuichi Shoji, Keiichi Imato

    International Conference on BioSensors, BioElectronics, BioMedical Devices, BioMEMS/NEMS & Applications (Bio4Apps 2017) 

    Presentation date: 2017.12

  • 温度応答性高分子ブラシを修飾した微細構造基板による細胞分離

    長瀬健一, 宿輪理紗, 小沼隆大, 大和雅之, 武田直也, 岡野光夫

    第39回日本バイオマテリアル学会大会 

    Presentation date: 2017.11

  • セルロースナノファイバー・チキソトロピックゲルでの三次元培養による長大な筋管形成

    米田憲司, 今任景一, 尹 棟鉉, 関口哲志, 庄子習一, 武田直也

    第39回日本バイオマテリアル学会大会 

    Presentation date: 2017.11

  • 導電性高分子をコアにもつ二層ファイバー足場を用いた電圧無印加での効率的筋管形成

    田中啓太, 中谷美沙, 今任景一, 武田直也

    第39回日本バイオマテリアル学会大会 

    Presentation date: 2017.11

  • 機械特性に優れる二成分系ファイバー基材への温度応答性ブラシ修飾と細胞分離

    宿輪理紗, 長瀬健一, 高橋宏信, 武田直也, 岡野光夫

    第39回日本バイオマテリアル学会大会 

    Presentation date: 2017.11

  • 導電層と細胞接着層の芯鞘二層マイクロファイバー足場における電位無負荷での効率的な筋組織形成誘導

    武田直也, 田中啓太, 中谷美沙, 今任景一

    第26回ポリマー材料フォーラム 

    Presentation date: 2017.11

  • チキソトロピー性セルロースゲルを用いた筋管形成を誘導するファイバー培養場の構築

    米田憲司, 今任景一, 尹 棟鉉, 関口哲志, 庄子習一, 武田直也

    第7回CSJ化学フェスタ2017 ―日本化学会秋季事業― 

    Presentation date: 2017.10

  • 導電層と細胞接着層の芯鞘二層マイクロファイバーの開発と筋組織誘導培養足場材料への応用

    田中啓太, 中谷美沙, 今任景一, 武田直也

    第7回CSJ化学フェスタ2017 ―日本化学会秋季事業― 

    Presentation date: 2017.10

  • 電荷移動相互作用で自己修復する室温駆動性エラストマーの設計

    中島英和, 今任景一, 武田直也

    第7回CSJ化学フェスタ2017 ―日本化学会秋季事業― 

    Presentation date: 2017.10

  • ラベルフリー細胞分離基材としての機械特性に優れた温度応答性マイクロファイバーの開発

    宿輪理紗, 桜田燿一, 長瀬健一, 高橋宏信, 武田直也, 岡野光夫

    第7回CSJ化学フェスタ2017 ―日本化学会秋季事業― 

    Presentation date: 2017.10

  • 細胞接脱着能を制御する光駆動型高分子ナノ薄膜の基底部界面の分子設計

    山本光哉, 今任景一, 武田直也

    第7回CSJ化学フェスタ2017 ―日本化学会秋季事業― 

    Presentation date: 2017.10

  • ポーラスマイクロファイバー基材を用いた灌流培養での流れストレス負荷による効率的な筋管形成誘導

    丸山 将夫, 尾崎 愛, 武田 直也

    第7回CSJ化学フェスタ2017 ―日本化学会秋季事業― 

    Presentation date: 2017.10

  • 紙とマイクロファイバーの重層基材を基盤とした自律駆動型灌流培養システムによる組織構築

    武田直也, 尾﨑 愛, 丸山将夫, 高橋宏信, 清水達也

    第55回日本人工臓器学会大会 

    Presentation date: 2017.09

  • 筋管組織の形成を効率的に誘導する導電層と細胞接着層の芯鞘二重マイクロファイバーからなる細胞培養材料の開発

    田中啓太, 中谷美沙, 今任景一, 武田直也

    第66回高分子討論会 

    Presentation date: 2017.09

  • 電荷移動相互作用を基盤とした生体温度で機能する自己修復性高分子材料の設計

    中島英和, 今任景一, 武田直也

    第66回高分子討論会 

    Presentation date: 2017.09

  • チキソトロピー性セルロースゲルを培養場とした多層ファイバー中での筋管形成誘導

    今任景一, 米田憲司, 田中 智, 武田直也

    第66回高分子討論会 

    Presentation date: 2017.09

  • ミクロ相分離構造の光スイッチングにより細胞挙動制御を実現するソフト界面の創製

    今任景一, 増田健一, 武田直也

    第46回医用高分子シンポジウム 

    Presentation date: 2017.07

  • Surface analysis of photo-responsive polymeric culture substrate showing contrary wettability changes to the polarity changes in isomerization of spiropyran

    Presentation date: 2017.05

  • Mechanistic study and controlling cell behavior of photo-transmutable microphase-separated structures in soft interfaces

    Presentation date: 2017.05

  • Effective induction of myotube formation using the self-driven perfusion culture system with porous polymeric fibers

    Presentation date: 2017.05

  • コラーゲンゲル接着による重層扁平 上皮細胞シートのEMT挙動解析

    葛西 善行, 武田 直也, 高木 亮, 大和 雅之

    第16回日本再生医療学会 

    Presentation date: 2017.03

  • メカノバイオロジー応用に向けた高延伸性培養基材と新規培養デバイスの作製

    舘野響樹, 秋山義勝, 坂口勝久, 高橋宏信, 清水達也, 武田直也

    第16回日本再生医療学会 

    Presentation date: 2017.03

  • 血管導入三次元組織の作製に向けたゲルファイバー包埋血管内皮細胞と細胞シートの積層化共培養

    近澤朋亮, 坂口勝久, 尹棟鉉, 関口哲志, 庄子習一, 清水達也, 武田直也

    第16回日本再生医療学会 

    Presentation date: 2017.03

  • 紙を基盤とした自律駆動型灌流培養システムによる血管内皮細胞のネットワーク形成誘導

    尾﨑 愛, 高橋宏信, 清水達也, 武田直也

    インテリジェント材料・システムシンポジウム2017 

    Presentation date: 2017.01

  • Reversible Controls of Wettability and Cell Adhesion of Photoresponsive Surface via Dynamic Conformational Change of Spiropyran-conjugated Block Copolymer

    Naoya TAKEDA, Di HE

    3rd International Conference on Biomaterials Science in Tokyo (ICBS2016)  (Tokyo) 

    Presentation date: 2016.11

  • 可逆的スイッチング機能を有するソフト界面の外部刺激応答機構と細胞接着挙動の解析

    増田健一, 武田直也

    日本バイオマテリアル学会シンポジウム2016 

    Presentation date: 2016.11

  • 紙を基盤とした自律駆動灌流培養のメカノストレス負荷による血管形成誘導

    尾﨑 愛, 高橋宏信, 清水達也, 武田直也

    日本バイオマテリアル学会シンポジウム2016 

    Presentation date: 2016.11

  • 温度応答性ナノインプリント微細構造表面での細胞接着・脱着挙動と細胞分離への応用

    宿輪理紗, 長瀬健一, 小沼隆大, 大和雅之, 武田直也, 岡野光夫

    日本バイオマテリアル学会シンポジウム2016 

    Presentation date: 2016.11

  • 光と熱によるミクロ相分離構造の可逆的スイッチングを実現する高分子界面設計と動的転換機構解析

    増田健一, 武田直也

    第6回CSJ化学フェスタ2016 

    Presentation date: 2016.11

  • 血管網構築に向けた効率的にメカノストレスを負荷する紙を基盤とした自律駆動型灌流培養システム

    尾﨑 愛, 高橋宏信, 清水達也, 武田直也

    第6回CSJ化学フェスタ2016 

    Presentation date: 2016.11

  • EBリソグラフパターニング基板での形態操作によるアストロサイトの遺伝子発現誘導

    町田奈穂, 武田直也

    第6回CSJ化学フェスタ2016 

    Presentation date: 2016.11

  • スピロピランを導入した同軸二層マイクロファイバーの分子設計と光刺激による細胞接着制御

    長田和歩, 武田直也

    第6回CSJ化学フェスタ2016 

    Presentation date: 2016.11

  • 生分解性高分子からなるナノポーラス・マイクロファイバー足場への細胞接着挙動解析

    田中啓太, 小町駿介, 長田和歩, 丸山将夫, 武田直也

    第6回CSJ化学フェスタ2016 

    Presentation date: 2016.11

  • 温度応答性ナノインプリント微細構造表面を用いた細胞分離システムの構築

    宿輪理紗, 長瀬健一, 小沼隆大, 大和雅之, 武田直也, 岡野光夫

    第6回CSJ化学フェスタ2016 

    Presentation date: 2016.11

  • ナノポーラス構造をもつ生分解性高分子マイクロファイバー 足場を用いた効率的な細胞培養

    田中啓太, 小町駿 介, 丸山将夫, 武田直也

    第65回高分子討論会 

    Presentation date: 2016.09

  • ナノインプリント微細構造を有する温度応答性表面の創製と細胞分離への応用

    宿輪 理紗, 長瀬 健一, 小沼 隆大, 大和 雅之, 武田 直也, 岡野 光夫

    第65回高分子討論会 

    Presentation date: 2016.09

  • Hybrid cell separation system by combining nanoimprint microstructure and thermoresponsive polymer surface

    Lisa SHUKUWA, Kenichi NAGASE, Takahiro ONUMA, Masayuki YAMATO, Naoya, Naoya TAKEDA, Teruo OKANO

    2rd International Symposium on Nanoarchitectonics for Mechanobiology 

    Presentation date: 2016.07

  • Self-driven Perfusion Culture System Using the Double-layered Scaffold of Paper and Microfibers for Effectively Applying Shear Stress to a Large Scale Cell Culture

    Ai OZAKI, Hironobu TAKAHASHI, Tatsuya SHIMIZU, Naoya TAKEDA

    2rd International Symposium on Nanoarchitectonics for Mechanobiology 

    Presentation date: 2016.07

  • 光応答性ポリマーをシース層に用いた同軸二層マイクロファイバー足場の作製と細胞接着制御

    長田和歩, 榑林哲也, 武田直也

    第65回高分子学会年次大会 

    Presentation date: 2016.05

  • Dynamic behaviors of spiropyran-introduced photo-responsive soft interface enabling reversible and long-term sustained cell patterning

    TAKEDA, Naoya, HE, Di, ARISAKA, Yoshinori

    Presentation date: 2016.03

  • 血管組織構築に向けた紙とマイクロファイバーの重層化基材を用いた自律駆動式灌流培養

    尾﨑 愛, 有坂慶紀, 高橋宏信, 清水達也, 武田直也

    第25回インテリジェント材料・システムシンポジウム 

    Presentation date: 2016.01

  • 光応答性ソフト界面におけるミクロ相分離構造転換の動的挙動解析とバイオ応用

    増田健一, 武田直也

    第25回インテリジェント材料・システムシンポジウム 

    Presentation date: 2016.01

  • Micro/Nano Fiber Scaffolds for Assembling Cells and Fabricating the Engineered Three-Dimensional Tissues

    Naoya Takeda  [Invited]

    International Conference on Bioelectronics, Biosensors, BioMedical Devices, BioMEMS/NEMS and Applications 2015 (Bio4Apps 2015).  (Fukuoka) 

    Presentation date: 2015.12

  • 紙とマイクロファイバーの重層化基材からなる自律駆動型灌流培養システムの開発

    尾﨑 愛, 有坂慶紀, 武田直也

    第37回バイオマテリアル学会 

    Presentation date: 2015.11

  • 相分離構造の光スイッチングにより細胞接・脱着制御を行うナノ薄膜培養基材の創製

    増田健一, 有坂慶紀, 武田直也

    第37回バイオマテリアル学会 

    Presentation date: 2015.11

  • 紙とマイクロファイバーの重層化基材を基盤とした自立駆動型灌流培養系の開発

    尾﨑 愛, 三澤博史, 有坂慶紀, 武田直也

    第5回CSJ化学フェスタ2015 

    Presentation date: 2015.10

  • ミクロ相分離構造の光スイッチング機能を持つソフト界面の創製と動的な細胞挙動制御

    増田健一, 有坂慶紀, 武田直也

    第5回CSJ化学フェスタ2015 

    Presentation date: 2015.10

  • 再生血管組織構築に向けた細胞接着性sheath層を持つ同軸二層ゲルファイバー培養場の作製

    後藤耀諒, 近澤朋亮, 南齋浩樹, 米田憲司, 有吉理紗, 尹 棟鉉, 関口哲志, 庄子習一, 武田直也

    第5回CSJ化学フェスタ2015 

    Presentation date: 2015.10

  • 表面修飾光応答性ブロック共重合体のダイナミックな高次構造変化による細胞の可逆的接・脱着制御

    何 迪, 有坂慶紀, 武田直也

    第5回CSJ化学フェスタ2015 

    Presentation date: 2015.10

  • 電子線リソグラフ・ナノパターンでの培養によるメカノストレス非依存的な間葉系幹細胞の分化誘起

    猿渡未来, 坪倉彩, 武田直也

    第5回CSJ化学フェスタ2015 

    Presentation date: 2015.10

  • ナノ相分離表面の光誘起構造転換と細胞マニピュレーション

    増田健一, 有坂慶紀, 武田直也

    第44回医用高分子シンポジウム 

    Presentation date: 2015.07

  • Novel spiropyran-incorporated polymer surface photo-dynamically changing micro domain pattern

    Presentation date: 2015.05

  • Purification of Mesenchymal Stem Cell using Thermo-responsive Polymer Brush Possessing Cationic Moieties

    Presentation date: 2015.05

  • 温度応答性高分子修飾ファイバーを用いたラベルフリー細胞分離系の作製

    第14回日本再生医療学会総会 

    Presentation date: 2015.03

  • 上皮細胞シートのESD創面への生着と創傷治癒機構のex vivoモデルによる解析

    第14回日本再生医療学会総会 

    Presentation date: 2015.03

  • ナノインプリント・リソグラフィーによる微細構造を有する細胞接脱着表面の作製

    第14回日本再生医療学会総会 

    Presentation date: 2015.03

  • Fabrication of Blood Capillary from Endothelial Cells in the Coaxial Double Layered Gel Fiber Produced by the 3D Sheath Flow Microfluidic Device

    Naoya Takeda  [Invited]

    International Conference on Bioelectronics, Biosensors, BioMedical Devices, BioMEMS/NEMS and Applications 2014 (Bio4Apps 2014)  (Shanghai) 

    Presentation date: 2014.11

  • Reversible photo-regulation of structural transition and cell adhesion on the functional surfaces modified with diblock copolymer containing spiropyran group

    Presentation date: 2014.09

  • 伸展可能な温度応答性細胞培養基材の物性評価と細胞シート剥離への応用

    第63回高分子討論会 

    Presentation date: 2014.09

  • 伸縮による温度応答型弾性表面の物性変化と細胞接着・脱着の制御

    第43回医用高分子シンポジウム 

    Presentation date: 2014.07

  • 配向化低密度「弦状」マイクロファイバー足場上で形成した高細胞密度筋管組織の機能構造解析

    第12回日本再生医療学会総会 

    Presentation date: 2013.03

  • エレクトロスピニング過程で化学架橋を施した配向化低密度「弦状」コラーゲンマイクロファイバー足場の作製と高細胞密度筋管組織の構築

    日本化学会第93春季年会(2013) 

    Presentation date: 2013.03

  • Differentiation of Mesenchymal Stem Cell on the Single Line Pattern in Nano-Size Fabricated on the Electron Beam Reactive Mask Material

    2nd International Conference of Biomaterials Science in Tsukuba (ICBS2013) 

    Presentation date: 2013.03

  • Fabrication of 3D Bundle of Muscle Fibers from Myoblasts Densely Filling the Microfibrous ‘String’ Scaffold; Studies on Structure and Actuation

    2nd International Conference of Biomaterials Science in Tsukuba (ICBS2013) 

    Presentation date: 2013.03

  • 細胞分離を目的とした温度応答性高分子ブラシ表面の系統的分子構造変化と機能評価

    日本バイオマテリアル学会大会シンポジウム2012 

    Presentation date: 2012.11

  • 弦状コラーゲンマイクロファイバー培養足場を用いた初代筋芽細胞からの収縮能をもつ三次元筋管組織構築

    日本バイオマテリアル学会大会シンポジウム2012 

    Presentation date: 2012.11

  • ナノパターンでの培養によるアストロサイトの星状形態誘導と遺伝子発現操作の検討

    日本バイオマテリアル学会大会シンポジウム2012 

    Presentation date: 2012.11

  • 紡糸中架橋化反応により作製した弦状コラーゲンマイクロファイバー培養足場の物性評価と配向筋管形成

    日本バイオマテリアル学会大会シンポジウム2012 

    Presentation date: 2012.11

  • Initiating Differentiation of Stem Cell into Neural Cell on the Nano Grooved Single Line Pattern Fabricated by Electron Beam Lithography

    Naoya Takeda

    Neuroscience 2012 

    Presentation date: 2012.10

  • Thermoresponsive Copolymer Brush Surface Incorporating Hydrophobic Unit for Mutimodal Cell Separating System

    9th international gel symposium 

    Presentation date: 2012.10

  • Cell Separating Thermoresponsive Polymer Brush Surfaces Prepared Through Surface-initiated ATRP

    9th international gel symposium 

    Presentation date: 2012.10

  • 三次元筋管組織形成誘導を実現する新規な「弦状」コラーゲンファイバー足場の開発

    第2回CSJ化学フェスタ 

    Presentation date: 2012.10

  • 表面マイクロパターンで誘起する一方向遊走のメカノストレスが幹細胞挙動へ与える効果

    第2回CSJ化学フェスタ 

    Presentation date: 2012.10

  • 電子線リソグラフ加工した高分子表面の微細構造によるヒト間葉系幹細胞の分化誘導

    第2回CSJ化学フェスタ 

    Presentation date: 2012.10

  • Development of Label-free Cell Separating System Based on The Functional Surface with Thermoresponsive Polymer Brush

    3rd TERMISWorld Congress 2012 "Tissue Engineering and Reenerative Medicine" 

    Presentation date: 2012.09

  • 電子線リソグラフ加工高分子レジスト基板を用いたナノ微細培養場での細胞の分化および機能操作

    第61回高分子討論会 

    Presentation date: 2012.09

  • The Novel String Scaffold of Aligned Collagen Microfiber Electrospun with Crosslinking Reaction

    第61回高分子討論会 

    Presentation date: 2012.09

  • 三次元筋管束を形成する「弦状」エレクトロスピン・マイクロファイバー細胞培養足場の開発

    平成24年度繊維学会秋季研究発表会 

    Presentation date: 2012.09

  • Cell Separating Surfaces Using Thermoresponsive Hydrophobized Copolymer Brush

    2nd International Symposium of Materials on Regenerative Medicine (2012 ISOMRM) 

    Presentation date: 2012.08

  • 配向化させた「弦状」エレクトロスピン・コラーゲンファイバー足場での効率的な三次元筋管形成

    第3回ソフトインターフェースの分子科学ワークショップ 「ソフト界面と計測・センシング」 

    Presentation date: 2012.08

  • 温度応答性表面によるラベルフリー細胞分離システムの設計

    第11回日本再生医療学会総会 

    Presentation date: 2012.06

  • 疎水性基を有する温度応答性高分子ブラシ表面による多段階温度制御細胞分離システムへの応用

    第11回日本再生医療学会総会 

    Presentation date: 2012.06

  • 「弦状」に中空維持・配向化させたエレクトロスピン・コラーゲンファイバー足場での三次元筋管形成誘導

    第11回日本再生医療学会総会 

    Presentation date: 2012.06

  • Differentiation of Mesenchymal Stem Cells with Culturing on the Nano-Structured Biointerfaces Fabricated by Electron Beam Lithography

    Satomi Aoki, Kenichi Tamura, Naoya Takeda

    IACIS 2012 (International Association of Colloid and Interface Scientists Conference) 

    Presentation date: 2012.05

  • Guiding Directional Migration by the Micropatterned Surface and Its Effect on the Proliferation/Differentiation Behaviors of Mesenchymal Stem Cell

    Aya Tsubokura, Naoya Takeda

    IACIS 2012 (International Association of Colloid and Interface Scientists Conference) 

    Presentation date: 2012.05

  • 細胞分離機能の高度化に向けた疎水性基を有する温度応答性高分子ブラシ表面の構築

    第61回高分子学会年次大会 

    Presentation date: 2012.05

  • 弦状エレクトロスピン・コラーゲンファイバー足場での配向筋管束の形成

    第61回高分子学会年次大会 

    Presentation date: 2012.05

  • 配向化エレクトロスピン・コラーゲンファイバー弦足場での三次元筋管形成誘導

    第21回インテリジェント材料/システムシンポジウム 

    Presentation date: 2012.01

  • 温度応答性インテリジェント表面の 精密設計による細胞選別能の高度化

    第21回インテリジェント材料/システムシンポジウム 

    Presentation date: 2012.01

  • 電子線リソグラフ微細パターンでの一方向遊走制御が間葉系幹細胞に与える効果

    第33回日本バイオマテリアル学会大会 

    Presentation date: 2011.11

  • 微少送液でゲル内培養液を直接置換する新規三次元培養系での細胞挙動評価

    第33回日本バイオマテリアル学会大会 

    Presentation date: 2011.11

  • 電子線リソグラフ微細パターンへの接着・伸展による間葉系幹細胞の分化誘導

    第33回日本バイオマテリアル学会大会 

    Presentation date: 2011.11

  • 電子線リソグラフ加工マイクロパターンによる細胞の一方向遊走誘起

    第33回日本バイオマテリアル学会大会 

    Presentation date: 2011.11

  • 温度応答性インテリジェント表面を用いた細胞分離システムの設計

    第33回日本バイオマテリアル学会大会 

    Presentation date: 2011.11

  • ナノ微細加工パターン培養での液性因子を用いない間葉系幹細胞の分化の検討

    第1回CSJ化学フェスタ —2011世界化学年記念大会— 

    Presentation date: 2011.11

  • 電子線リソグラフ加工マイクロパターンによる細胞の一方向遊走誘起

    第1回CSJ化学フェスタ —2011世界化学年記念大会— 

    Presentation date: 2011.11

  • Micro/nano topography by electron beam lithography to manipulate cell migration and differantiation

    Presentation date: 2011.09

  • Preparation of Thermoresponsive Intelligent Interface for Cell Separation

    60th SPSJ Annual Meeting 

    Presentation date: 2011.05

  • Guiding long-range directional cell migration using newly developed micropatterned substrates

    The 91th Annual Meeting of CSJ 

    Presentation date: 2011.03

  • 一本の微細な細長い溝(トレンチ)状ナノパターンでの培養による間葉系幹細胞の分化の検討

    第10回 日本再生医療学会総会 

    Presentation date: 2011.03

  • Fabrication of cell patterning system with micro-nano resolution by electron beam lithography and adhesion of single cell on the nano structure

    Presentation date: 2011.01

  • Directing Cell Migration by Controlling Cell Shape with Microstructured Substrates

    Presentation date: 2011.01

  • 3D cell culture in collagen gel applied continuous solvent flow at small rate; Long-term culture and effect of the shear stress to the cells

    Presentation date: 2011.01

  • Navigation of neurite elongation and fabrication of the networks of neural cells with fine resolution on the novel cell culture surface patterned by electron beam lithography

    Neuro2010 

    Presentation date: 2010.09

▼display all

Specific Research

  • 高分子ファイバーを用いた生体ガス検出センサーの開発

    2021  

     View Summary

    生体ガス中の揮発性有機化合物の一つであるエタノールガスを光学的に検出する高感度センサーの開発を行った。微細なファイバーのメッシュを担持体として、ここに検出を担う分子と補因子を組み込んでセンサーを作製した。光学的に検出する装置も作製した。比表面積の大きい微細なファイバー間隙をガスが透過するため、効率的なガス検出が可能となった。さまざまな材料を担持体としたセンサーを作製してガス検出機能を定量的に解析し、材料の最適化を行った。センサーの応答速度も解析し、既存のセンサーとの比較において優位性を示した。

  • セルロースナノファイバーの外科手術材料への応用

    2020  

     View Summary

    外科手術に用いる材料としての、セルロースナノファイバーの有効性を評価した。さまざまな形状や分子構造を有する複数種類のセルロースナノファイバーについて物理化学特性を詳細に定量測定し、外科材料として適当な種類と使用条件の最適化を行った。また、手術を模擬した実験において、使用目的を十分に満たす性能を有することを確認し、既存の代替材料と比較しても高い機能が期待できることを実証した。さらに、使用前後で材料特性が大きく変化することを新たに見出し、その機構についても解析を行った。さらに、生物学的手法をもちいて、セルロースナノファイバーが生体に与える影響についても解析を行った。

  • マイクロファイバーの複合材料化によるバイオ基材への応用

    2020  

     View Summary

    電界紡糸マイクロファイバーに他の材料や分子を組み合わせて複合化し、新たなバイオ材料を作製した。ナノ構造を導入した二次元材料と組み合わせた複合材料は、細胞培養基材として有効であることを実証した。培養した細胞は高次構造を有する組織を形成し、生体組織と同様の機能を発現することも確認した。電界紡糸マイクロファイバーに他の生体分子を組み合わせた複合材料については、ファイバーに用いる材料の最適化を行った。その結果、生体分子の機能を維持したまま複合化できることを実証した。さらに、ファイバーを加工してデバイス構築へと展開し、ファイバー形状がデバイス性能に影響することも見出した。

  • 気液界面培養システムによる生体の構造・機能を有する腸管上皮モデル組織の作製

    2019  

     View Summary

    医薬品や機能性食品等の開発に有用な、生体同様の形態と機能を有する腸管上皮モデル組織を構築した。腸管の内腔は気層に面するため、本研究では、気相に接した気液界面で腸管上皮細胞を培養できる、紙の上に高分子マイクロファイバーをマット状に紡糸した二層構造からなる培養器材を開発した。上層の高分子ファイバーで高い細胞接着性を実現し、下層の紙に保持した培養液をファイバーの間隙から上層へ供給して気液界面培養を可能とした。作製した腸管上皮モデル組織は、小腸に特有の絨毛突起様の三次元構造体を形成し、apical面には微絨毛や細胞間には密着結合が形成された。また、消化酵素や薬物代謝酵素も発現し、粘液産生能が顕著に増大した。

  • 紙とマイクロファイバーの重層基材を基盤とした再生生体組織培養システムの創製

    2017   今任 景一

     View Summary

     本研究では、紙の上にエレクトロスピニング法で生分解性高分子のマイクロファイバーを紡糸した二層基材を作製した。マイクロファイバー層には細胞接着と培養場の機能を担わせ、特にファイバー表面にBreath Figure法でナノサイズの多孔を形成させて細胞接着性の向上を実現した。さらに、紙の吸水能とサイフォン原理を利用して培養液を自律的に灌流するシステムとし、筋芽細胞を播種して細胞の挙動に及ぼす培養液の流れストレスの効果を定量的に評価した。その結果、静置培養時と比較して細胞増殖ならびに筋管形成の増大が引き起こされることが分かった。すなわち、流れストレスは筋管形成への効率的な分化刺激となることが示された。

  • 再生医工学への応用に向けた芯/鞘二重構造マイクロファイバーの高機能化戦略

    2017   今任 景一

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    &nbsp; エレクトロスピニング法により芯/鞘二重構造を持つ機能性マイクロファイバーを作製し、細胞培養基材に応用した。紫外光照射でナノ鞘層の高分子表面のぬれ性が増すファイバー基材では、光刺激により細胞の接着性が有意に増大した。この光制御機能は、光応答性を担う分子の高分子鎖内への導入率が高いほど向上した。また、芯層に導電性高分子を導入して、細胞接着性の高分子の鞘層で被覆したファイバー材料も作製した。このファイバー基材上で筋芽細胞を培養すると、電位を負荷しない条件にもかかわらず鞘層のみの足場と比較して増殖率および筋管形成率が有意に増大し、再生筋組織の作製基材として優れることを見出した。

  • 界面π造形の可逆的光制御システムの構築と細胞挙動操作

    2017  

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     光応答性スピロピラン分子(SP)を導入した疎水ブロックと親水ブロックからなるジブロック共重合体を合成し、アルキル基で疎水化したガラス基材へナノ薄膜状に修飾した光応答性基板を開発した。特に、高分子組成および基材界面のアルキル鎖を系統的に変化させ、光刺激に対する表面物性ならびに細胞接着性の変化を評価した。その結果、表面親疎水性変化(親水→疎水)がSPの光異性化に伴う極性変化(疎水→親水)と相反し、また紫外光の照射で細胞が接着・伸展し可視光の照射では細胞は脱着した。この現象は、高分子鎖の動的な高次構造変化に起因し、高分子間のみならず高分子薄膜と基材界面での相互作用も重要であることを明らかにした。

  • 紙とマイクロファイバーの重層基材からなる自律駆動型灌流培養システムによる組織構築

    2016  

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     本研究では、紙とゼラチンファイバーからなる重層化細胞培養足場を基盤とし、紙の毛細管現象を培養液の自律的な駆動力に利用すると共にサイフォン原理を組み合わせて連続的な送液が可能である、これまでにないシンプルで大面積かつ開放系での細胞培養場を有する新たな自律駆動型の灌流培養システムを開発した。さらに、血管内皮細胞単独ならびに血管内皮細胞と異種細胞との灌流共培養を行った。その結果、静置培養との比較において、血管内皮細胞が流れに応答して挙動を顕著に変化させることを見出した。これより、本灌流培養システムが、血管組織構築に有効なシェアストレスを効果的に負荷できることを示した。

  • 同軸重層マイクロゲルファイバーを基盤とした成熟血管組織作製システムの創成

    2015  

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    細胞を組み上げて三次元の生体組織を作製する再生医療研究において、拡散により栄養や酸素が十分に行き届くのが困難な組織内部へこれらを血流により供給する長大な毛細血管組織の構築は極めて重要である。本研究では、基本ユニットを積層化させて作製し、三次元的な層流を安定して形成できる新たなマイクロ流体デバイスを開発し、血管内皮細胞からの血管組織構築に有用な同軸二層のマイクロゲルファイバー培養足場の作製。新デバイスによりゲルファイバーの作製効率の向上を達成すると共に、アルギン酸に細胞接着ペプチドを化学修飾した新たなゲル材料の作製も進めた。Fabrication of large, long, and/or thick engineeredtissues, which are implantable to human body for regenerative medicine,attracts great interest. For this purpose, fabrication and introduction of 3Dand long blood vessels to maintain cells in the internal area of 3D tissues arecritical and the challenging subjects. In this study, the co-axial doublelayered microgel fibers (micrometers in diameter but tens of centimeters inlength) were prepared as the 3D culture scaffolds. The 3D sheath flowmicrofluidic device was improved and the new-type device, which was composed thestacking basic units, was developed. Effective fabrication of the microgel fiberswas also achieved using this device. Vascular endothelial cells were three-dimensionallycultured in the core layer of the microgel fiber and formed luminal structure. Thegel materials increased in cell-adhesion worked to enhance formation of luminalcapillary.

  • 配向化コラーゲン線維束三次元足場の傾斜的石灰化と腱骨移行部再生組織構築への応用

    2014   有坂 慶紀

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    腱組織は、配向したⅠ型コラーゲン線維が傾斜的に石灰化しながら線維軟骨組織を経て骨組織へと埋入する。腱断裂において線維軟骨は再生されないために、再生組織の作製が期待されている。本研究では、生体組織構造を模倣して、配向Ⅰ型コラーゲンマイクロファイバーに傾斜的にハイドロキシアパタイトを析出させた新たな三次元培養足場を構築した。さらに、この足場上で培養した線維芽細胞と間葉系幹細胞が、足場構造に応じて生体内と同様な細胞形態へと変化することを見出した。さらに、傾斜的石灰化構造に応じた間葉系幹細胞の腱細胞への分化誘導を示唆する新たな知見を得た。The tendon tissue consists of uniformly oriented typeI collagen fibers. These collagen fibers gradually bear hydroxyapatite and areembedded into bone tissue. For constructing a regenerative tendon tissue, a three-dimensionaland oriented collagen microfiber scaffold, on which hydroxyapatite gradually deposited,was fabricated by mimicking the native tissue. Depending on the graduallychanging components of the scaffold, the fibroblasts and mesenchymal stem cells(MSCs) cultured on the scaffold was found to change their shape as they take invivo. Furthermore, the novel data suggesting that MSCs differentiated intotendon cells depending on the scaffold structure was obtained.

  • マイクロ材料工学を基盤とした大型三次元生体組織の構築と複合組織化による機能制御

    2014   庄子 習一, 関口 哲志, 有坂 慶紀, 奥 仁美, 小町 駿介, 南齋 浩樹, 後藤 耀諒, 近澤 朋亮

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    再生医療応用を目指し、細胞を組み上げて三次元の生体組織を作製する研究が大きな注目を集めている。管腔化した三次元組織である毛細血管は、他の再生組織の深部へ血流を供給する応用も期待され、重要な研究のターゲットである。本研究では、血管内皮細胞から長大な毛細血管組織の作製を目的とし、長さ数十cm ~ mスケール、直径約100μmの同軸二層のCore[芯]–Sheath[鞘]型(CS型)ゲルファイバー三次元培養場を、独自に開発した三次元的な層流を形成できるマイクロ流体デバイスにより作製した。Core層に血管内皮細胞を懸濁して導入し、Sheath層に生体適合性の高い混合ゲルを用いて包埋培養し、管腔化した毛細血管組織の作製に成功した。Fabricating engineered tissues by assembling individual cells is ofgreat interest to engineers and scientists. Tissues and organs of a human bodyhave three-dimensional fine structures, and their size is centimeter and/ormeter scale. Therefore, for regenerative medicine applications, engineeredtissues should be the same size, and it is necessary to develop innovativetechnologies to assemble the cells into a three-dimensional and largestructure. In this study, we developed the novel microfluidic device, which canform three-dimensional laminar flow in its micro channel, and we applied this deviceto fabricate the engineered blood capillaries (vessels).The three-dimensional laminar flow consists of a core layer and acoaxially surrounding sheath layer. By continuously introducing different solmaterials to each layer and subsequently gelating them, the double layeredhydrogels of fiber shape were fabricated. Vascular endothelial cells were alsosuccessively introduced in the core layer and three-dimensionally cultured inthe hydrogel fiber so that the cells autonomously developed into long bloodcapillary. Various biopolymers were used as gel materials and examined how theyaffect gel fiber formation and tissue development.

  • 分子・細胞・組織の階層的集積と統合による運動器官生体ロボットの創製

    2013  

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      生体には、細胞や基質が一方向に配向して構造化し機能を発現する組織が見られる。骨格筋組織では、筋芽脂肪が縦一列に整列・融合して筋管へと分化し、筋管は筋線維へと成熟する。多数の筋線維は同一方向に配向しながら三次元の束となって大きな収縮力を示す骨格筋組織を形成する。さらに、骨格筋の端は腱組織へと連なるが、この腱組織は主に配向したⅠ型コラーゲン線維からなり、線維軟骨へと移行しながら傾斜的に石灰化が進み、最終的に骨へ埋入する。このような構造と機能をもった骨格筋と腱の三次元組織をin vitroで工学的に作製することを目指し、エレクトロスピニング法により生体適合性材料を用いて作製したマイクロファイバーを基盤とした三次元培養足場を構築した。 骨格筋組織の作製では、ファイバー同士に適度な間隔を与えつつ同一方向に配向させて中空に張った「弦状」の足場を作製した。これまでに、I型コラーゲンで作製した長さ1 cm x 幅5 mmの弦状足場でラット初代骨格筋細胞を培養して三次元筋組織を構築した成果を踏まえて、より大型の組織作製に取り組んだ。このため、I型コラーゲンに生分解性高分子であるポリカプロラクトン(PCL)を組み合わせてファイバーの強度を向上させ、ドラムコレクタを用いてファイバーの配向化を効率的に行なうことで、長さ2 cm x 幅1 cmへと足場の大型化を達成した。この足場にラット初代骨格筋細胞を播種し21~24日間培養することで、拍動120回/分、収縮率2%程度の自発的収縮能を有し横紋構造をもつ骨格筋組織の形成を達成した。蛍光抗体染色と画像解析による組織中央部における横紋組織の存在率は、約70%と見積もられた。 腱組織については、長さ1 cmの弦状足場の半分を塩化カルシウム溶液とリン酸水素二ナトリウム溶液へ交互に浸漬し、ファイバー束の半分にHApを析出させた傾斜的石灰化三次元足場を作製した。XPSによる元素組成の解析から、傾斜的にHApが析出していることが確認された。この足場に線維芽細胞NIH3T3、前骨芽細胞MC3T3-E1さらにヒト骨髄由来間葉系幹細胞(hbmMSC)をそれぞれ播種したところ、傾斜化した足場の線維部、移行部、HAp化部の各局所において細胞の形態・分布が変化した。また、腱マーカーであるスクレラキシス(Scx)遺伝子の発現をqRT-PCRにより定量的に解析したところ、足場材料の違いによりhbmMSCsのScx遺伝子発現の変化が示唆された。 以上の結果を踏まえて、今後、各再生組織の構造と機能の改善をさらに進めると共に、別途作製を進めている神経束組織や血管組織とも組み合わせて複合組織・器官の構築を目指す。

  • 三次元可変集束層流マイクロ流体デバイスによるマクロ複合生体組織の創製

    2013  

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    [緒言]細胞からのin vitroでの生体組織の作製研究は近年大きな注目を集めており、特に、移植医療への応用も可能なセンチメートル以上の大型組織の作製が望まれている。長大な血管組織や神経組織を作製するためには、マイクロ径のファイバー状ハイドロゲル内で細胞を三次元包埋培養するアプローチが有力な手法と考え、本研究では、Core-Sheath状の同軸二層型ゲルファイバーを連続的に作製し得る新規なマイクロ流体デバイス(三次元可変シースフローマイクロ流体デバイス)を開発した。このゲルファイバー内部で血管内皮細胞または神経系細胞を包埋培養し、血管組織や神経組織の作成に向けた内皮細胞の融合および神経突起の伸長挙動を評価すると共に、これらを効率的に誘導する生体高分子ゲル材料についても検討した。[結果と考察]三次元可変シースフローマイクロ流体デバイスは、細胞を流すCoreとなる層に外側の流路からゲル作製用高分子溶液(Sheath層)を二段階であてて周囲を取り囲みながらCore層を流路中央へと寄せることで、Core-Sheath状の同軸二層を実現する設計とした。さらに、最外層の流路から供給したゲル化水溶液で高分子をゲル化させ、流れに乗せてゲルファイバーをデバイス外へと排出させる工夫を施した。まず、それぞれの流路を流れる各溶液の流量比を調節することによって、Core層の直径を10 μmから40 μm程度まで調節できることを確認した。続いて、Ca2+との錯形成でハイドロゲルを形成するアルギン酸をSheath層から導入し、最外流路からはCa2+を供給して細胞をCore層に包埋したゲルファイバーを作製した。Core-Sheath状の同軸二層型ゲルファイバーの形成は、細胞と一緒にCore層にFITC標識コラーゲンを導入して蛍光でCore層のみを可視化することで確認した。Core層にPC12細胞を導入し、Sheath層のゲル材料にはアルギン酸のみを用いた実験では、三日間の培養において神経様突起がCore層に沿って伸長する様子が観察され、また80%以上のviabilityが得られた。さらに、より細胞培養に適した培養場とするため、Sheath層のハイドロゲル材料について検討を加えた。多糖であるアルギン酸に同じく多糖類のカラギーナンを組み合わせたり、細胞の三次元培養足場に頻用されるゼラチンやフィブリン(いずれもタンパク質)とアルギン酸との組み合わせについても評価した。ウシ大動脈血管内皮細胞(BAEC)をCore層に導入して検討したところ、カラギーナンとの組み合わせではゲルが脆くなってしまったのに対し、ゼラチンおよびフィブリンと組み合わせたゲルファイバーでは十分なゲル強度が確保され、90%以上のviabilityが得られた。さらに細胞が一列に連なった構造体が観察された。今後のさらなる研究進展により、本デバイスを用いて作製したゼラチンやフィブリンを主体とするゲルファイバーを用いて、長大な毛細血管や神経組織の構築が期待される。

  • 神経筋接合部により運動機能を制御する三次元再生骨格筋組織の構築

    2012  

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    骨格筋組織の発生では、一列に連なった筋芽細胞が融合して多核の筋管となり、筋管は横紋構造を示す収縮能をもった筋線維へと成熟する。さらに筋線維は、コラーゲンに富んだ筋内膜に覆われて同一方向へ配向した筋束を形成する。本研究では、I型コラーゲンを用いて独自にデザインした「弦状」の細胞培養足場を構築すると共に、細胞密度が高く収縮能をもつ大きな三次元の骨格筋組織の構築を進め、その構造と機能を解析した。 エレクトロスピニング法で紡糸中にグルタルアルデヒド(GA)架橋を施しながら、I型コラーゲン・マイクロファイバーを同一方向に配向させつつ、足場材料密度を下げて細胞が入り込む適度な間隔を保って中空に張り、三次元の「弦状」足場を作製した。ファイバーの配向化のため、2 cmの間隔を空けた平行な2枚の銅板をエレクトロスピニングのターゲット板に用いた。架橋により機械的強度の向上(無架橋との比較でヤング率と引張り強度共に、約2.7倍に増大)と耐水溶性が付与された。また、コラーゲン溶液(HFP溶液)の粘度、GAとの混合比(コラーゲン:GA = 1 : 0.2 (w/w))、紡糸時間(30秒)など諸条件の最適化により、平均ファイバー径は2 &#61549;m、平均ファイバー間隔は6 &#61549;mに制御された。足場に播種されたマウス筋芽細胞C2C12細胞およびラット初代筋芽細胞は、ファイバー間の空隙に入り込みながらファイバーに沿って配向・融合して筋管を形成し、これら筋管は同一方向へ高密度に集合して全長1 cmの三次元細胞構造体を形成した。三次元筋線維束組織の断面および切片をそれぞれSEMおよびワンギーソン染色法により画像解析したところ、断面のサイズは40 x 120 &#61549;mで、設計通りにコラーゲンファイバーは組織全体に散在していた。 代表的な骨格筋マーカーであるミオシン重鎖で蛍光免疫染色を行い観察したところ、筋管組織の平均長は290 &#61549;mであり一部の筋構造体には筋線維に特徴的な横紋がみられた。そこで、C2C12細胞から作製した三次元筋線維束組織に電気刺激を与えたところ、収縮が見られなかった。一方で、初代筋芽細胞から作製した組織では電気刺激を与えなくても自発的な収縮が観察され、運動機能を有する再生骨格筋組織の構築を達成した。今後は電気刺激与えてそのパターンにより運動を制御することを試みる。また、平行して取り組んでいる初代神経細胞から作製する再生神経束組織と共培養し、神経筋接合部の形成を評価すると共に、神経組織からのシグナル入力による骨格筋組織の運動制御にも取り組む予定である。

  • 自律神経組織体による心筋細胞組織体の拍動機能制御システムの構築

    2009  

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    1. 研究目的神経-心筋の共培養や生体情報伝達解析ならびに細胞機能制御のプラットフォームとなる培養システムの構築を目指し、その基盤技術としての、神経系細胞(初代神経細胞、神経モデル細胞、グリア細胞)の単一細胞を精緻に配置して、神経細胞ネットワークを作製する培養システムの構築に注力した。この神経系細胞の単一細胞ネットワーク構築には、ナノスケールの半導体加工に汎用されている高分子の電子線レジストを用いたリソグラフィー技術を応用した。ナノサイズのパターンを描画した電子線レジストは、半導体加工ではマスク材として用いられた後に除去されるが、本研究ではこのナノ描画高分子表面を除去することなく直接に細胞のパターニングが可能な培養基板へと応用した。2. 実験電子線照射部位のみに最小解像度20 nmで精緻なパターンを作製可能なpoly[1-chloro-(methyl acrylate)-co-(1-methylstylene)]を主体とした高分子ポジ型レジストをガラス表面に400 nm厚で塗布し、電子線照射および現像処理により一辺20~50 μmの正方形パターンをアレイ状に作製した。未照射部位はpluronicF108で修飾をして、細胞の接着を妨げた。これにより、アレイ状の細胞接着領域に対して、神経細胞、アストロサイト、ならびに神経モデル細胞であるPC12細胞が単一細胞レベルで接着が可能とした。更に、100 nm幅の微細なライン状のパターンに沿って、ナノサイズである神経突起の伸長方向を精確に誘導することを試みた。3. 結果と考察PluronicF108の表面修飾は有効に機能し、高いコントラストをもって、電子線で描画したパターン上のみへ細胞が接着した。一辺20 μmの正方形の細胞接着領域をアレイ状に並べたパターンでは、アストロサイトとPC12細胞を用いた場合に、各パターンに単一の細胞を配置できた。また、蛍光染色を用いた解析では、正方形パターンの縁の各辺に沿ってアストロサイト細胞のアクチンフィラメントの集積がみられ、細胞が細胞接着領域のパターンを正確に認識して接着斑を形成していることが強く示唆された。さらに、アストロサイトやPC12細胞の細胞突起構造を、100 nm幅の微細なラインパターンに沿って精確に伸展させることも達成した。胎仔ラット由来の初代神経細胞についても、単一細胞レベルでのアストロサイトとの共培養を実現した。これらの結果から、本単一細胞培養システムは、パターン形状の適切なデザインにより、任意の形状での精緻な細胞ネットワーク形成や神経回路形成への応用が可能であることを示した。

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Syllabus

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Committee Memberships

  • 2015
    -
    Now

    日本バイオマテリアル学会  評議員

  • 2013
    -
    Now

    Bio4Apps  Program Committee member

  • 2002
    -
    Now

    (一財)マイクロマシンセンター  国内外技術動向調査委員会委員

  • 2010
    -
    2018

    未踏科学技術協会 インテリジェント・ナノ材料研究会  企画幹事

  • 2015
    -
     

    Japanese Society for Biomaterials  Councilor

  • 2006
    -
    2012

    Japanese Society for Biomaterials  Councilor

  • 2006
    -
    2012

    日本バイオマテリアル学会  評議員

  • 2010
    -
     

    Intelligent Nano Materials Society, The Society of Non-Traditional Technology  Member of Managing Committee

  • 2008.11
    -
     

    日本バイオマテリアル学会  日本バイオマテリアル学会シンポジウム2008 実行委員

  • 2008.10
    -
     

    日本コンピュータ外科学会  第17回 日本コンピュータ外科学会大会 実行委員

  • 2002
    -
     

    Micromachine Center  Member of the committee, Research Department

  • 2001.12
    -
     

    日本ME学会  第15回 日本ME学会秋季大会 実行委員

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Social Activities

  • マイクロ・ナノバイオテクノロジーを駆使して、血管や筋肉組織をつくる

    Harima Quarterly 2018, 137:6―9.  (137:6―9.) 

    2018
    -
     

Media Coverage

  • 筋芽細胞培養に新たな道 (報道)

    日経産業新聞(報道)  

    2017.11

  • 培養細胞、生存率6倍 (報道)

    日経産業新聞(報道)  

    2011.01