Updated on 2022/05/26

写真a

 
SAKAGUCHI, Katsuhisa
 
Affiliation
Faculty of Science and Engineering, Graduate School of Advanced Science and Engineering
Job title
Associate Professor(without tenure)
Mail Address
メールアドレス
Profile

早稲田大学 先進理工学研究科 生命理工学専攻 准教授
1998-2002年 早稲田大学 機械工学科 学部
2002-2004年 早稲田大学大学院 生命理工学専攻 修士課程
2006-2010年 早稲田大学大学院 生命理工学専攻 博士課程
2009-2012年 早稲田大学 総合機械工学科助手
2012-2015年 早稲田大学 理工学術院総合研究所 次席研究員/研究院講師
2015-2019年 早稲田大学 理工学術院総合研究所 主任研究員/研究院准教授
2019年~ 早稲田大学 先進理工学研究科 生命理工学専攻 准教授

Research Institute

  • 2020
    -
    2022

    理工学術院総合研究所   兼任研究員

Degree

  • Waseda University   Doctor of Philosophy in Engineering

Professional Memberships

  •  
     
     

    ライフサポート学会

  •  
     
     

    日本人工臓器学会

  •  
     
     

    日本機械学会

  •  
     
     

    日本再生医療学会

 

Research Areas

  • Robotics and intelligent system   Biomechanics

  • Biomedical engineering   Tissue Engineering

  • Biofunction and bioprocess engineering

Research Interests

  • 人間医工学

  • 医用機械工学

  • 細胞培養工学

  • 組織工学

  • 再生医療

Papers

  • 培養肉生産に向けたウシ筋芽細胞の攪拌浮遊培養

    坂口 勝久, 田中 龍一郎

    生物工学会誌   100 ( 4 ) 165 - 168  2022.04

    Authorship:Lead author, Corresponding author

    DOI

  • Cell-Based Microfluidic Device Utilizing Cell Sheet Technology

    Katsuhisa Sakaguchi, Kei Akimoto, Masanori Takaira, Ryu-ichiro Tanaka, Tatsuya Shimizu, Shinjiro Umezu

    Cyborg and Bionic Systems   2022   1 - 8  2022.01  [Refereed]

    Authorship:Lead author, Corresponding author

     View Summary

    The development of microelectromechanical systems has resulted in the rapid development of polydimethylpolysiloxane (PDMS) microfluidic devices for drug screening models. Various cell functions, such as the response of endothelial cells to fluids, have been elucidated using microfluidic devices. Additionally, organ-on-a-chip systems that include organs that are important for biological circulation, such as the heart, liver, pancreas, kidneys, and brain, have been developed. These organs realize the biological circulation system in a manner that cannot be reproduced by artificial organs; however, the flow channels between the organs are often artificially created by PDMS. In this study, we developed a microfluidic device consisting only of cells, by combining cell sheet technology with microtitanium wires. Microwires were placed between stacked fibroblast cell sheets, and the cell sheets adhered to each other, after which the microwires were removed leaving a luminal structure with a size approximately equal to the arteriolar size. The lumen structure was constructed using wires with diameters of 50, 100, 150, and 200 <italic>μ</italic>m, which were approximations of the arteriole diameters. Furthermore, using a perfusion device, we successfully perfused the luminal structure created inside the cell sheets. The results revealed that a culture solution can be supplied to a cell sheet with a very high cell density. The biofabrication technology proposed in this study can contribute to the development of organ-on-a-chip systems.

    DOI

  • Perfusable vascular tree like construction in 3D cell-dense tissues using artificial vascular bed

    Yusuke Tobe, Jun Homma, Katsuhisa Sakaguchi, Hidekazu Sekine, Kiyotaka Iwasaki, Tatsuya Shimizu

    Microvascular Research   141   104321  2022.01  [Refereed]

    Authorship:Corresponding author

    DOI

  • In vitro ballooned hepatocytes can be produced by primary human hepatocytes and hepatic stellate cell sheets

    Nobuhiro Hasui, Katsuhisa Sakaguchi, Tetsuya Ogawa, Yoshihiro Sakamoto, Tatsuya Shimizu

    Scientific Reports   12 ( 1 )  2021.12  [Refereed]

    Authorship:Corresponding author

     View Summary

    Abstract

    Despite the increasing prevalence of Nonalcoholic steatohepatitis (NASH) worldwide, there is no effective treatment available for this disease. “Ballooned hepatocyte” is a characteristic finding in NASH and is correlated with disease prognosis, but their mechanisms of action are poorly understood; furthermore, neither animal nor in vitro models of NASH have been able to adequately represent ballooned hepatocytes. Herein, we engineered cell sheets to develop a new in vitro model of ballooned hepatocytes. Primary human hepatocytes (PHH) and Hepatic stellate cells (HSC) were co-cultured to produce cell sheets, which were cultured in glucose and lipid containing medium, following which histological and functional analyses were performed. Histological findings showed hepatocyte ballooning, accumulation of fat droplets, abnormal cytokeratin arrangement, and the presence of Mallory–Denk bodies and abnormal organelles. These findings are similar to those of ballooned hepatocytes in human NASH. Functional analysis showed elevated levels of TGFβ-1, SHH, and p62, but not TNF-α, IL-8. Exposure of PHH/HSC sheets to a glucolipotoxicity environment induces ballooned hepatocyte without inflammation. Moreover, fibrosis is an important mechanism underlying ballooned hepatocytes and could be the basis for the development of a new in vitro NASH model with ballooned hepatocytes.

    DOI

  • Bioengineering of a scaffold-less three-dimensional tissue using net mould

    Katsuhisa Sakaguchi, Yusuke Tobe, Jiayue Yang, Ryu-ichiro Tanaka, Kumiko Yamanaka, Jiro Ono, Tatsuya Shimizu

    Biofabrication    2021.09  [Refereed]

    Authorship:Lead author, Corresponding author

    DOI

  • Perfusable System Using Porous Collagen Gel Scaffold Actively Provides Fresh Culture Media to a Cultured 3D Tissue.

    Chikahiro Imashiro, Kai Yamasaki, Ryu-Ichiro Tanaka, Yusuke Tobe, Katsuhisa Sakaguchi, Tatsuya Shimizu

    International journal of molecular sciences   22 ( 13 )  2021.06  [Refereed]  [International journal]

    Authorship:Corresponding author

     View Summary

    Culturing three-dimensional (3D) tissues with an appropriate microenvironment is a critical and fundamental technology in broad areas of cutting-edge bioengineering research. In addition, many technologies have engineered tissue functions. However, an effective system for transporting nutrients, waste, or oxygen to affect the functions of cell tissues has not been reported. In this study, we introduce a novel system that employs diffusion and convection to enhance transportation. To demonstrate the concept of the proposed system, three layers of normal human dermal fibroblast cell sheets are used as a model tissue, which is cultured on a general dish or porous collagen scaffold with perfusable channels for three days with and without the perfusion of culture media in the scaffold. The results show that the viability of the cell tissue was improved by the developed system. Furthermore, glucose consumption, lactate production, and oxygen transport to the tissues were increased, which might improve the viability of tissues. However, mechanical stress in the proposed system did not cause damage or unintentional functional changes in the cultured tissue. We believe that the introduced culturing system potentially suggests a novel standard for 3D cell cultures.

    DOI PubMed

  • 実験室で培養される人工組織・臓器

    坂口勝久

    生物工学会誌   99 ( 4 ) 197 - 197  2021.04  [Invited]

    Authorship:Lead author, Corresponding author

    DOI

  • Functional Analysis of Induced Human Ballooned Hepatocytes in a Cell Sheet-Based Three Dimensional Model

    Botao Gao, Katsuhisa Sakaguchi, Tetsuya Ogawa, Yuki Kagawa, Hirotsugu Kubo, Tatsuya Shimizu

    Tissue Engineering and Regenerative Medicine    2021.01  [Refereed]

    DOI

  • Reconstruction of a Vascular Bed with Perfusable Blood Vessels Using a Decellularized Porcine Small Intestine for Clinical Application

    Yusuke Tobe, Katsuhisa Sakaguchi, Jun Homma, Kazunori Sano, Eiji Kobayashi, Hidekazu Sekine, Kiyotaka Iwasaki, Tatsuya Shimizu, Mitsuo Umezu

    IFMBE Proceedings     284 - 292  2021  [Refereed]

    Authorship:Corresponding author

    DOI

  • Trends in Tissue Engineering and Cell Sheet Engineering

    Materials Science and Technology of Japan   57 ( 5 ) 172 - 175  2020.10  [Refereed]  [Invited]

    Authorship:Lead author, Corresponding author

  • Measuring the Contractile Force of Multilayered Human Cardiac Cell Sheets.

    Katsuhisa Sakaguchi, Hiroaki Takahashi, Yusuke Tobe, Daisuke Sasaki, Katsuhisa Matsuura, Kiyotaka Iwasaki, Tatsuya Shimizu, Mitsuo Umezu

    Tissue engineering. Part C, Methods   26 ( 9 ) 485 - 492  2020.09  [Refereed]  [International journal]

    Authorship:Lead author

     View Summary

    Three-dimensional (3D) cardiac tissue reconstruction using tissue engineering technology is a rapidly growing area of regenerative medicine and drug screening development. However, there remains an urgent need for the development of a method capable of accurately measuring the contractile force of physiologically relevant 3D myocardial tissues to facilitate the prediction of human heart tissue drug sensitivity. To this end, our laboratory has developed a novel drug screening model that measures the contractile force of cardiac cell sheets prepared using temperature-responsive culture dishes. To circumvent the difficulties that commonly arise during the stacking of cardiomyocyte sheets, we established a stacking method using centrifugal force, making it possible to measure 3D myocardial tissue. Human induced pluripotent stem cell-derived cardiomyocytes were seeded in a temperature-responsive culture dish and processed into a sheet. The cardiac cell sheets were multilayered to construct 3D cardiac tissue. Measurement of the contractile force and cross-sectional area of the multilayered 3D cardiac tissue were then obtained and used to determine the relationship between the cross-sectional area of the cardiac tissue and its contractile force. The contractile force of the 1-, 3-, and 5-layer tissues increased linearly in proportion to the cross-sectional area. A result of 6.4 mN/mm2, accounting for one-seventh of the contractile force found in adult tissue, was obtained. However, with 7-layer tissues, there was a sudden drop in the contractile force, possibly because of limited oxygen and nutrient supply. In conclusion, we established a method wherein the thickness of the cell sheets was controlled through layering, thus enabling accurate evaluation of the cardiac contractile function. This method may enable comparisons with living heart tissue while providing information applicable to regenerative medicine and drug screening models.

    DOI PubMed

  • Printing support hydrogels for creating vascular-like structures in stacked cell sheets

    Ryu-ichiro Tanaka, Katsuhisa Sakaguchi, Shinjiro Umezu

    Artificial Life and Robotics   25 ( 2 ) 199 - 203  2020.05  [Refereed]

    DOI

  • Microfluidic vascular-bed devices for vascularized 3D tissue engineering: tissue engineering on a chip

    Hiroaki Takehara, Katsuhisa Sakaguchi, Hidekazu Sekine, Teruo Okano, Tatsuya Shimizu

    Biomedical Microdevices   22 ( 1 ) 9 - 9  2020.03  [Refereed]  [International journal]

     View Summary

    In this report, we describe a microfluidic vascular-bed (micro-VB) device providing a platform for 3D tissue engineering with vascular network formation. The micro-VB device allows functional connections between endothelial capillaries of heterogeneous sections (5-100 μm in diameter) and artificial plastic tubes or reservoirs (1-10 mm in diameter). Moreover, the micro-VB device can be installed in a standard 100 mm-diameter Petri dish. Endothelial networks in 3D engineered tissues were obtained by cellular self-assembly on the device, after co-culturing of human umbilical vein endothelial cells (HUVECs) and normal human dermal fibroblasts (NHDFs) in fibrin gel. Endothelial capillary connection between vascularized tissues and microfluidic channels, mimicking arteries and veins, was confirmed by perfusion of fluorescent microspheres. The micro-VB devices were compatible with the use of commercially available culture dishes and did not require the involvement of additional equipment. Thus, these micro-VB devices are expected to substantially improve the routine application of 3D tissue engineering to regenerative medicine.

    DOI PubMed

  • In Vitro Production of Human Ballooned Hepatocytes in a Cell Sheet-based Three-dimensional Model.

    Botao Gao, Katsuhisa Sakaguchi, Katsuhisa Matsuura, Tetsuya Ogawa, Yuki Kagawa, Hirotsugu Kubo, Tatsuya Shimizu

    Tissue engineering. Part A   26 ( 1-2 ) 93 - 101  2020.01  [Refereed]  [International journal]

     View Summary

    Ballooned hepatocytes (BH) are enlarged, abnormal hepatocytes, which are usually involved in liver diseases, in particular, nonalcoholic steatohepatitis (NASH). However, formation of BHs in vitro has been seldom reported. This study reported an in vitro strategy to produce human BHs in a cell sheet-based three-dimensional (3D) model where primary human hepatocytes were cocultured with normal human dermal fibroblasts. Enlargement of hepatocytes (2.3 times larger than normal, p < 0.01), loss of cytoplasmic keratin, appearance of Mallory-Denk bodies (MDBs), and abundant fat droplets accumulation were observed after only a few days culture. Additionally, ultrastructural characteristic findings of BHs in human NASH, including enlarged mitochondria with crystalline inclusions, dilated endoplasmic reticulum, and MDBs formation were also observed in the 3D model. Furthermore, pathophysiological features of human NASH, such as increased secretion of sonic hedgehog ligands and myofibroblast activation were found. This study reports in vitro production of human BHs by using a cell sheet-based 3D model. Similar histological, ultrastructural, and pathophysiological features to human NASH are discovered in this model. This model may facilitate study of BHs and increase our knowledge of the pathogenesis of human liver diseases. Impact Statement Human ballooned hepatocytes (BH), which are present in nonalcoholic steatohepatitis (NASH) are mainly studied based on human liver biopsies and animal models. In this study, human BHs can be successfully reproduced in a cell sheet-based in vitro model, which, as far as we know, is the first in vitro model that recapitulates so many histological and ultrastructural hallmarks of BHs found in human NASH. Additionally, this study also demonstrated presence of some NASH pathophysiological features. This model may facilitate the study of hepatocellular ballooning and prove beneficial in translational preclinical drug discovery in NASH.

    DOI PubMed

  • Adequate taylor couette flow-mediated shear stress is useful for dissociating human iPS cell-derived cell aggregates.

    Katsuhisa Matsuura, Masanori Wada, Katsuhisa Sakaguchi, Yuki Matsuhashi, Tatsuya Shimizu

    Regenerative therapy   12   6 - 13  2019.12  [Refereed]  [International journal]

     View Summary

    Pluripotent stem cell including induced pluripotent stem cells (iPSC) are promising cell sources for regenerative medicine and for three-dimensional suspension culture technologies which may enable the generation of robust numbers of desired cells through cell aggregation. Although manual procedure is widely used for dissociating cell aggregates, the development of non-manual procedures using devices will contribute to efficient cell manufacturing. In the present study, we developed novel cell aggregate dissociation devices with a rotating cylinder inside based on taylor couette flow-mediated shear stress. The shear stress can be increased according to an increase in the size of the rotating cylinder inside the devices and the rotation rate. Adequate device size and suitable rotation rate efficiently dissociated cell aggregates after the undifferentiated expansion and the cardiac differentiation of human iPSC. These finding suggest that non-manual device procedure might be useful for harvesting single cells from human iPSC-derived cell aggregates.

    DOI PubMed

  • Pulsatile tubular cardiac tissues fabricated by wrapping human iPS cells-derived cardiomyocyte sheets.

    Shinpei Tsuruyama, Katsuhisa Matsuura, Katsuhisa Sakaguchi, Tatsuya Shimizu

    Regenerative therapy   11   297 - 305  2019.12  [Refereed]  [International journal]

     View Summary

    The purpose of this study was to fabricate pulsatile tubular cardiac tissue using cell sheet based-tissue engineering. First, we fabricated human induced pluripotent stem cell (hiPSc)-derived cardiomyocyte sheets and normal human dermal fibroblast (NHDF) sheets which are harvested from temperature responsive culture dishes only by lowering the temperature. Then tubular cardiac tissues are formed by wrapping one hiPSc-derived cardiomyocyte sheet and three NHDF sheets around an octagonal column, and both ends of the tubular tissue were covered with fibrin and collagen gel. The octagonal column with the tubular tissue was connected to an in vitro circulation system in a culture box. After four-day culture, the cardiac tissue survived and pulsated spontaneously in the circulation system. Furthermore, the analysis with a Millar catheter inserted into the cardiac tubes revealed significant inner pressure changes generated by their beating. In addition, the tubular cardiac tissue pulsated in response to the electrical stimulation. Although histological analyses demonstrated that cardiac troponin T-positive cells stratified the inner surface of the tubular tissues, gene expression analyses showed an immature state of these cardiomyocytes. Thus, cell sheet-based tissue engineering realized human pulsatile tubular cardiac tissue fabrication and we believe that these tubular cardiac tissues should contribute to future drug screening and regenerative therapy for heart diseases.

    DOI PubMed

  • Simple action potential measurement of cardiac cell sheet utilizing electronic sheet

    Takashi Ohya, Kazuki Nakazono, Tetsutaro Kikuchi, Daisuke Sasaki, Katsuhisa Sakaguchi, Tatsuya Shimizu, Kenjiro Fukuda, Takao Someya, Shinjiro Umezu

    Artificial Life and Robotics   23 ( 3 ) 321 - 327  2018.09  [Refereed]

    DOI

  • Low-temperature culturing improves survival rate of tissue-engineered cardiac cell sheets

    Katsuhisa Sakaguchi, Yuto Hinata, Yuki Kagawa, Kiyotaka Iwasaki, Satoshi Tsuneda, Tatsuya Shimizu, Mitsuo Umezu

    Biochemistry and Biophysics Reports   14   89 - 97  2018.07  [Refereed]

    Authorship:Lead author

    DOI

  • Elucidation of the Relationship Between the Contractile Force and the Cross-sectional Area of Stacked human iPSC-derived Cardiomyocyte Sheets

    Hiroaki Takahashi, Katsuhisa Sakaguchi, Yusuke Tobe, Kazuki Nakazono, Daisuke Sasaki, Katsuhisa Matsuura, Tatsuya Sshimizu, Mitsuo Umezu

    The Proceedings of the Bioengineering Conference Annual Meeting of BED/JSME   2018.30 ( 0 )  2018  [Refereed]

    DOI

  • Fundamental characteristics of printed gelatin utilizing micro 3D printer

    Ryu-ichiro Tanaka, Katsuhisa Sakaguchi, Shinjiro Umezu

    Artificial Life and Robotics   22 ( 3 ) 316 - 320  2017.09  [Refereed]

     View Summary

    Gelatin is useful for biofabrication, because it can be used for cell scaffolds and it has unique properties. Therefore, we attempted to fabricate biodevices of gelatin utilizing micro 3D printer which is able to print with high precision. However, it has been difficult to fabricate 3D structure of gelatin utilizing 3D printer, because a printed gelatin droplet on the metal plate electrode would spread before solidification. To clear this problem, we developed a new experimental set-up with a peltier device that can control temperature of the impact point. At an impact point temperature of 80 °C, the spreading of printed gelatin droplets was prevented. Therefore, we were able to print a ball gelatin. In addition, we were able to print a narrower gelatin line than at an impact point temperature of 20 °C.

    DOI

  • The potential of cell sheet technique on the development of hepatocellular carcinoma in rat models

    Alaa T. Alshareeda, Katsuhisa Sakaguchi, Mohammed Abumaree, Nur Khatijah Mohd Zin, Tatsuya Shimizu

    PLOS ONE   12 ( 8 ) e0184004  2017.08  [Refereed]

     View Summary

    Background
    Hepatocellular carcinoma (HCC) is considered the 3rd leading cause of death by cancer worldwide with the majority of patients were diagnosed in the late stages. Currently, there is no effective therapy. The selection of an animal model that mimics human cancer is essential for the identification of prognostic/predictive markers, candidate genes underlying cancer induction and the examination of factors that may influence the response of cancers to therapeutic agents and regimens. In this study, we developed a HCC nude rat models using cell sheet and examined the effect of human stromal cells (SCs) on the development of the HCC model and on different liver parameters such as albumin and urea.
    Methods
    Transplanted cell sheet for HCC rat models was fabricated using thermo-responsive culture dishes. The effect of human umbilical cord mesenchymal stromal cells (UC-MSCs) and human bone marrow mesenchymal stromal cells (BM-MSCs) on the developed tumour was tested. Furthermore, development of tumour and detection of the liver parameter was studied. Additionally, angiogenesis assay was performed using Matrigel.
    Results
    HepG2 cells requires five days to form a complete cell sheet while HepG2 co-cultured with UC-MSCs or BM-MSCs took only three days. The tumour developed within 4 weeks after transplantation of the HCC sheet on the liver of nude rats. Both UC-MSCs and BM-MSCs improved the secretion of liver parameters by increasing the secretion of albumin and urea. Comparatively, the UC-MSCs were more effective than BM-MSCs, but unlike BM-MSCs, UC-MSCs prevented liver tumour formation and the tube formation of HCC.
    Conclusions
    Since this is a novel study to induce liver tumour in rats using hepatocellular carcinoma sheet and stromal cells, the data obtained suggest that cell sheet is a fast and easy technique to develop HCC models as well as UC-MSCs have therapeutic potential for liver diseases. Additionally, the data procured indicates that stromal cells enhanced the fabrication of HepG2 cell sheets. This provides the foundation for future research using stromal cells in preclinical and clinical investigations.

    DOI PubMed

  • Fabrication of micro-gelatin fiber utilizing coacervation method

    Takafumi Arai, Ryuichiro Tanaka, Katsuhisa Sakaguchi, Shinjiro Umezu

    Artificial Life and Robotics   22 ( 2 ) 197 - 202  2017.06  [Refereed]

     View Summary

    Biotechnology has drastically been advanced by the development of iPS and ES cells, which are representative forms induced pluripotent stem cells. In the micro/nano bio field, the development of cells and Taylor-made medicine for a potential treatment of incurable diseases has been a center of attention. The melting point of gelatin is between 25 and 33 °C, and the sol–gel transition occurs in low temperature. This makes the deformation of this useful biomaterial easy. The examples of gelatin fiber applications are suture threads, blood vessel prosthesis, cell-growth-based materials, filter materials, and many others. Because the cell size differs depending on the species and applications, it is essential to fabricate gelatin fibers of different diameters. In this paper, we have developed a fabrication method for gelatin fibers the coacervation method. We fabricated narrow gelatin fibers having a diameter over 10 μm.

    DOI

  • Fabrication of micro-alginate gel tubes utilizing micro-gelatin fibers

    Katsuhisa Sakaguchi, Takafumi Arai, Tatsuya Shimizu, Shinjiro Umezu

    Japanese Journal of Applied Physics   56 ( 5 )  2017.05  [Refereed]

    Authorship:Lead author

     View Summary

    Tissues engineered utilizing biofabrication techniques have recently been the focus of much attention, because these bioengineered tissues have great potential to improve the quality of life of patients with various hard-to-treat diseases. Most tissues contain micro-tubular structures including blood vessels, lymphatic vessels, and bile canaliculus. Therefore, we bioengineered a micro diameter tube using alginate gel to coat the core gelatin gel. Micro-gelatin fibers were fabricated by the coacervation method and then coated with a very thin alginate gel layer by dipping. A micro diameter alginate tube was produced by dissolving the core gelatin gel. Consequently, these procedures led to the formation of micro-alginate gel tubes of various shapes and sizes. This biofabrication technique should contribute to tissue engineering research fields. (C) 2017 The Japan Society of Applied Physics

    DOI

  • Optical coherence microscopy of living cells and bioengineered tissue dynamics in high-resolution cross-section

    Akiyuki Hasegawa, Yuji Haraguchi, Hirotoshi Oikaze, Yasuhiro Kabetani, Katsuhisa Sakaguchi, Tatsuya Shimizu

    Journal of Biomedical Materials Research Part B - Applied Biomaterials   105 ( 3 ) 481 - 488  2017.04  [Refereed]

     View Summary

    Optical coherence tomography (OCT) is a valuable tool in the cross-sectional observation/analysis of three-dimensional (3-D) biological tissues, and that histological observation is important clinically. However, the resolution of the technology is approximately 10-20 m. In this study, optical coherence microscopy (OCM), a tomographic system combining OCT technology with a microscopic technique, was constructed for observing cells individually with a resolution at the submicrometer level. Cells and 3-D tissues fabricated by cell sheet technology were observed by OCM. Importantly, the cell nuclei and cytoplasm could be clearly distinguished, and the time-dependent dynamics of cell-sheet tissues could be observed in detail. Additionally, the 3-D migration of cells in the bioengineered tissue was also detected using OCM and metal-labeled cells. Bovine aortic endothelial cells, but not NIH3T3 murine embryonic skin fibroblasts, actively migrated within the 3-D tissues. This study showed that the OCM system would be a valuable tool in the fields of cell biology, tissue engineering, and regenerative medicine. (c) 2015 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 105B: 481-488, 2017.

    DOI PubMed

  • Vascularized Tissue-Engineered Model for Studying Drug Resistance in Neuroblastoma

    Aranzazu Villasante, Kakatsuhisa Sakaguchi, Jinho Kim, Nai-Kong Cheung, Masamichii Nakayama, Hasam Parsa, Teruo Okano, Tatsuya Shimizu, Gordana Vunjak-Novakovic

    Theranostics   7 ( 17 ) 4099 - 4117  2017  [Refereed]

    DOI

  • Thicker three-dimensional tissue from a "symbiotic recycling system" combining mammalian cells and algae

    Yuji Haraguchi, Yuki Kagawa, Katsuhisa Sakaguchi, Katsuhisa Matsuura, Tatsuya Shimizu, Teruo Okano

    Scientific Reports   7   41594  2017.01  [Refereed]

     View Summary

    In this paper, we report an in vitro co-culture system that combines mammalian cells and algae, Chlorococcum littorale, to create a three-dimensional (3-D) tissue. While the C2C12 mouse myoblasts and rat cardiac cells consumed oxygen actively, intense oxygen production was accounted for by the algae even in the co-culture system. Although cell metabolism within thicker cardiac celllayered tissues showed anaerobic respiration, the introduction of innovative co-cultivation partially changed the metabolism to aerobic respiration. Moreover, the amount of glucose consumption and lactate production in the cardiac tissues and the amount of ammonia in the culture media decreased significantly when co-cultivated with algae. In the cardiac tissues devoid of algae, delamination was observed histologically, and the release of creatine kinase (CK) from the tissues showed severe cardiac cell damage. On the other hand, the layered cell tissues with algae were observed to be in a good histological condition, with less than one-fifth decline in CK release. The co-cultivation with algae improved the culture condition of the thicker tissues, resulting in the formation of 160 mu m-thick cardiac tissues. Thus, the present study proposes the possibility of creating an in vitro "symbiotic recycling system" composed of mammalian cells and algae.

    DOI PubMed

  • Controlling sheer stress in a suspension culture using couette flow for efficient proliferation of HEK 293 cells

    Nur Khatijah Mohd Zin, Katsuhisa Sakaguchi, Yuji Haraguchi, Takanobu Yagi, Tatsuya Shimizu, Mitsuo Umezu

    Fluid Mechanics Open Access   3 ( 1 )  2016.05  [Refereed]

    Authorship:Corresponding author

    DOI

  • 細胞シート技術を用いた立体組織再生

    坂口 勝久

    日本画像学会誌   55 ( 1 ) 83 - 87  2016.02

    Authorship:Lead author, Corresponding author

    DOI

  • Controlling shape and position of vascular formation in engineered tissues by arbitrary assembly of endothelial cells

    Hiroaki Takehara, Katsuhisa Sakaguchi, Masatoshi Kuroda, Megumi Muraoka, Kazuyoshi Itoga, Teruo Okano, Tatsuya Shimizu

    Biofabrication   7 ( 4 ) 045006  2015.12  [Refereed]

     View Summary

    Cellular self-assembly based on cell-to-cell communication is a well-known tissue organizing process in living bodies. Hence, integrating cellular self-assembly processes into tissue engineering is a promising approach to fabricate well-organized functional tissues. In this research, we investigated the capability of endothelial cells (ECs) to control shape and position of vascular formation using arbitral-assembling techniques in three-dimensional engineered tissues. To quantify the degree of migration of ECs in endothelial network formation, image correlation analysis was conducted. Positive correlation between the original positions of arbitrarily assembled ECs and the positions of formed endothelial networks indicated the potential for controlling shape and position of vascular formations in engineered tissues. To demonstrate the feasibility of controlling vascular formations, engineered tissues with vascular networks in triangle and circle patterns were made. The technique reported here employs cellular self-assembly for tissue engineering and is expected to provide fundamental beneficial methods to supply various functional tissues for drug screening and regenerative medicine.

    DOI PubMed

  • Time Course of Cell Sheet Adhesion to Porcine Heart Tissue after Transplantation

    Dehua Chang, Tatsuya Shimizu, Yuji Haraguchi, Shuai Gao, Katsuhisa Sakaguchi, Mitsuo Umezu, Masayuki Yamato, Zhongmin Liu, Teruo Okano

    PLoS One   10 ( 10 ) e0137494  2015.10  [Refereed]

     View Summary

    Multilayered cell sheets have been produced from bone marrow-derived mesenchymal stem cells (MSCs) for investigating their adhesion properties onto native porcine heart tissue. Once MSCs reached confluence after a 7-day culture on a temperature-responsive culture dish, a MSCs monolayer spontaneously detached itself from the dish, when the culture temperature was reduced from 37 to 20 degrees C. The basal extracellular matrix (ECM) proteins of the single cell sheet are preserved, because this technique requires no proteolytic enzymes for harvesting cell sheet, which become a basic building block for assembling a multilayer cell sheet. The thickness of multilayered cell sheets made from three MSC sheets was found to be approximately 60 mu m. For investigating the adhesion properties of the basal and apical sides, the multilayered cell sheets were transplanted onto the surface of the heart's left ventricle. Multilayered cell sheets were histological investigated at 15, 30, 45 and 60 minutes after transplantation by hematoxylin eosin (HE) and azan dyes to determine required time for the adhesion of the multilayered sheets following cell-sheet transplantation. The results showed that only the basal side of multilayered cell sheets significantly enhanced the sheets adhesion onto the surface of heart 30 minutes after transplantation. This study concluded that (1) cell sheets had to be transplanted with its basal side onto the surface of heart tissue and (2) at least 30 minutes were necessary for obtaining the histological adhesion of the sheets to the heart tissue. This study provided clinical evidence and parameters for the successful application of MSC sheets to the myocardium and allowed cell sheet technology to be adapted clinical cell-therapy for myocardial diseases.

    DOI PubMed

  • 3D組織構築のためのバイオリアクタの開発

    坂口勝久, 清水達也

    人工臓器   44 ( 1 ) 57 - 61  2015.06

    Authorship:Lead author

    DOI

  • Construction of three-dimensional vascularized cardiac tissue with cell sheet engineering

    Katsuhisa Sakaguchi, Tatsuya Shimizu, Teruo Okano

    Journal of Controlled Release   205   83 - 88  2015.05  [Refereed]

    Authorship:Lead author

     View Summary

    Construction of three-dimensional (3D) tissues with pre-isolated cells is a promising achievement for novel medicine and drug-discovery research. Our laboratory constructs 3D tissues with an innovative and unique method for layering multiple cell sheets. Cell sheets maintain a high-efficiently regenerating function, because of the higher cell density and higher transplantation efficiency, compared to other cell-delivery methods. Cell sheets have already been applied in clinical applications for regenerative medicine in treating patients with various diseases. Therefore, in our search to develop a more efficient treatment with cell sheets, we are constructing 3D tissues by layering cell sheets. Native animal tissues and organs have an abundance of capillaries to supply oxygen and nutrients, and to remove waste molecules. In our investigation of vascularized cardiac cell sheets, we have found that endothelial cells within cell sheets spontaneously form blood vessel networks as in vivo capillaries. To construct even thicker 3D tissues by layering multiple cell sheets, it is critical to have a medium or blood flow within the vascular networks of the cell sheets. Therefore, to perfuse medium or blood in the cell sheet vascular network to maintain the viability of all cells, we developed two types of vascular beds; (1) a femoral muscle-based vascular bed, and (2) a synthetic collagen gel-based vascular bed. Both vascular beds successfully provide the critical flow of culture medium, which allows 12-layer cell sheets to survive. Such bioreactor systems, when combined with cell sheet engineering techniques, have produced functional vascularized 3D tissues. Here we explain and discuss the various processes to obtain vascular networks by properly connecting cell sheets and the engineering of 3D tissues. (C) 2014 Elsevier B.V. All rights reserved.

    DOI PubMed

  • In vitro engineering of vascularized tissue surrogates.

    Katsuhisa Sakaguchi, Tatsuya Shimizu, Shigeto Horaguchi, Hidekazu Sekine, Masayuki Yamato, Mitsuo Umezu, Teruo Okano

    Scientific reports   3   1316 - 1316  2013.02  [Refereed]  [International journal]

    Authorship:Lead author

     View Summary

    In vitro scaling up of bioengineered tissues is known to be limited by diffusion issues, specifically a lack of vasculature. Here, we report a new strategy for preserving cell viability in three-dimensional tissues using cell sheet technology and a perfusion bioreactor having collagen-based microchannels. When triple-layer cardiac cell sheets are incubated within this bioreactor, endothelial cells in the cell sheets migrate to vascularize in the collagen gel, and finally connect with the microchannels. Medium readily flows into the cell sheets through the microchannels and the newly developed capillaries, while the cardiac construct shows simultaneous beating. When additional triple-layer cell sheets are repeatedly layered, new multi-layer construct spontaneously integrates and the resulting construct becomes a vascularized thick tissue. These results confirmed our method to fabricate in vitro vascularized tissue surrogates that overcomes engineered-tissue thickness limitations. The surrogates promise new therapies for damaged organs as well as new in vitro tissue models.

    DOI PubMed

  • In vitro fabrication of functional three-dimensional tissues with perfusable blood vessels.

    Hidekazu Sekine, Tatsuya Shimizu, Katsuhisa Sakaguchi, Izumi Dobashi, Masanori Wada, Masayuki Yamato, Eiji Kobayashi, Mitsuo Umezu, Teruo Okano

    Nature communications   4   1399 - 1399  2013.01  [Refereed]  [International journal]

     View Summary

    In vitro fabrication of functional vascularized three-dimensional tissues has been a long-standing objective in the field of tissue engineering. Here we report a technique to engineer cardiac tissues with perfusable blood vessels in vitro. Using resected tissue with a connectable artery and vein as a vascular bed, we overlay triple-layer cardiac cell sheets produced from coculture with endothelial cells, and support the tissue construct with media perfused in a bioreactor. We show that endothelial cells connect to capillaries in the vascular bed and form tubular lumens, creating in vitro perfusable blood vessels in the cardiac cell sheets. Thicker engineered tissues can be produced in vitro by overlaying additional triple-layer cell sheets. The vascularized cardiac tissues beat and can be transplanted with blood vessel anastomoses. This technique may create new opportunities for in vitro tissue engineering and has potential therapeutic applications.

    DOI PubMed

  • Fabrication of functional three-dimensional tissues by stacking cell sheets in vitro.

    Yuji Haraguchi, Tatsuya Shimizu, Tadashi Sasagawa, Hidekazu Sekine, Katsuhisa Sakaguchi, Tetsutaro Kikuchi, Waki Sekine, Sachiko Sekiya, Masayuki Yamato, Mitsuo Umezu, Teruo Okano

    Nature protocols   7 ( 5 ) 850 - 8  2012.04  [Refereed]  [International journal]

     View Summary

    The fabrication of 3D tissues retaining the original functions of tissues/organs in vitro is crucial for optimal tissue engineering and regenerative medicine. The fabrication of 3D tissues also contributes to the establishment of in vitro tissue/organ models for drug screening. Our laboratory has developed a fabrication system for functional 3D tissues by stacking cell sheets of confluent cultured cells detached from a temperature-responsive culture dish. Here we describe the protocols for the fabrication of 3D tissues by cell sheet engineering. Three-dimensional cardiac tissues fabricated by stacking cardiac cell sheets pulsate spontaneously, synchronously and macroscopically. Via this protocol, it is also possible to fabricate other tissues, such as 3D tissue including capillary-like prevascular networks, from endothelial cells sandwiched between layered cell sheets. Cell sheet stacking technology promises to provide in vitro tissue/organ models and more effective therapies for curing tissue/organ failures.

    DOI PubMed

  • Novel device for transplantation of cell sheet and evaluation of thin polymer films by atomic force microscopy

    Ryohei Takeuchi, Kazuhiro Fukumori, Katsuhisa Sakaguchi, Yutaka Terajima, Tatsuya Shimizu, Teruo Okano, Mitsuo Umezu

    International Symposium on Micro-NanoMechatronics and Human Science     168 - 173  2011.11  [Refereed]

     View Summary

    Cell therapy is expected to a new tool to treat refractory diseases. In heart regeneration, it has been firstly conducted with needle injection of cell suspensions. Recently, cell sheet engineering emerged as another method of cell therapy. Cell sheet is prepared with a temperature responsive dish by temperature reduction. It is a thin-patch-like tissue construct and its thickness is several tens of micrometers. It is composed of cells and intrinsic extra cellular matrix only. The transplantation of the cell sheet has been already conducted in animal experiments and even in clinical trials. The cell sheet is transplanted at the surface of the heart, but it is difficult to transplant the cell sheet under the beating heart. To overcome this difficulty, we designed a device that was composed of two thin polymer films that have different friction. The films were made of polyurethane, polyethylene, or polypropylene. The cell sheet was set up on the device by sandwiching it with the less frictional film and the more frictional film. In this paper, using two different films having the different friction, the cell sheet was successfully transplanted to the static round polymer surface, the harvested heart, and even the beating heart of pig by removing the films step by step using the difference in friction. Also, surface properties such as friction, adhesion force and roughness of the films were studied by an atomic force microscopy (AFM). From the results of the study, the friction of the film was found to be likely proportional to the adhesion force and the inverse of roughness. © 2011 IEEE.

    DOI

  • Fabrication of Three-Dimensional Tissues with Perfused Microchannels

    Katsuhisa Sakaguchi, Tatsuya Shimizu, Kiyotaka Iwasaki, Masayuki Yamato, Mitsuo Umezu, Teruo Okano

    IFMBE Proceedings     1213 - 1216  2009.11  [Refereed]

    Authorship:Lead author

    DOI

  • ティッシュエンジニアリングによる心筋再生医療の現状と展望

    坂口勝久, 清水達也, 梅津光生

    東京女子医科大学雑誌   79 ( 3 ) 108 - 112  2009.09  [Refereed]

    Authorship:Lead author

  • Development of Bioreactor System for Bioengineering Three-Dimensional Myocardial Tissues

      21 ( 3 ) 104 - 109  2009  [Refereed]

    Authorship:Lead author

    DOI

▼display all

Misc

  • Transplantation of three-dimensional cardiac tissue by cell sheet rapid stacking

    田原滉大, 坂口勝久, 本間順, 松浦勝久, 岩崎清隆, 清水達也

    日本生物工学会大会講演要旨集   73rd  2021

    J-GLOBAL

  • 実験室で培養される人工組織・臓器

    坂口勝久

    生物工学会誌   99 ( 4 )  2021

    J-GLOBAL

  • Trends in Tissue Engineering and Cell Sheet Engineering

    坂口勝久

    材料の科学と工学   57 ( 5 ) 172 - 175  2020.10  [Refereed]  [Invited]

    J-GLOBAL

  • 移植可能な血管床の作製のための脱細胞化組織骨格の再細胞化手法の検討

    戸部友輔, 坂口勝久, 関根秀一, 本間順, 佐野和紀, 小林英司, 清水達也, 梅津光生

    日本再生医療学会総会(Web)   19th  2020.03

    J-GLOBAL

  • ネットモールド法により構築可能な3次元組織厚に関する基礎検討

    楊佳悦, 大野次郎, 坂口勝久, 戸部友輔, 清水達也, 梅津光生, 梅津光生

    日本再生医療学会総会(Web)   19th  2020

    J-GLOBAL

  • Investigation into the effect of ovarian hormone to the thickness of human endometrial stromal cell sheets in vitro.

    藤田真央, 坂口勝久, 藏本吾郎, 梅津信二郎, 清水達也

    日本機械学会関東支部・精密工学会・茨城大学工学部茨城講演会講演論文集(CD-ROM)   28th  2020

    J-GLOBAL

  • 細胞シートを積層化した立体肝組織モデルの機能評価

    森山千弘, 坂口勝久, 高博韜, 小川徹也, 加川友己, 久保寛嗣, 清水達也, 梅津光生

    日本再生医療学会総会(Web)   18th  2019.03

    J-GLOBAL

  • ヒトiPS細胞由来心筋細胞を用いたフィブリンを利用した高速スタンプ技術によるヒト立体心筋組織構築と移植

    中園一紀, 坂口勝久, 清水達也, 梅津光生

    日本再生医療学会総会(Web)   18th  2019.03

    J-GLOBAL

  • 重層化心筋細胞シートの移植治療実現のための脱細胞化ブタ小腸を利用したヒトに移植可能な血管床の開発

    戸部友輔, 坂口勝久, 佐野和紀, 関根秀一, 清水達也, 小林英司, 梅津光生

    日本再生医療学会総会(Web)   18th  2019.03

    J-GLOBAL

  • 子宮内膜組織構築に向けた灌流培養デバイスの開発・検討

    坂田周平, 坂口勝久, 鈴木崇, 藏元吾朗, 梅津信二郎, 清水達也

    日本再生医療学会総会(Web)   18th  2019.03

    J-GLOBAL

  • 脂肪由来間葉系幹細胞と血管内皮細胞の共培養による細胞シート内血管網付与の条件検討

    山崎快, 坂口勝久, 戸部友輔, 清水達也, 梅津光生

    日本再生医療学会総会(Web)   18th  2019.03

    J-GLOBAL

  • 灌流バイオリアクターを用いた血管網付立体組織の構築

    坂口勝久, 戸部友輔, 佐野和紀, 関根秀一, 松浦勝久, 小林英司, 清水達也, 梅津光生

    日本再生医療学会総会(Web)   18th  2019

    J-GLOBAL

  • ファイバ・プローブ型OCTの光干渉断層撮影装置(OCT)用いたブタ管状血管床内腔面の観察・評価

    小林真里, 岩名信一, 坂口勝久, 清水達也

    日本再生医療学会総会(Web)   18th  2019

    J-GLOBAL

  • 細胞シートを用いたin vitro下の子宮内膜組織三次元構築と受精卵着床・浸潤解明に関する研究

    鈴木崇, 藏本吾郎, 坂田周平, 高木惣一, 坂口勝久, 清水達也

    日本再生医療学会総会(Web)   18th  2019

    J-GLOBAL

  • 細胞シート積層立体肝組織モデルによる薬剤クリアランスの予測

    小川徹也, 加川友己, 森山千弘, 高博韜, 坂口勝久, 久保寛嗣, 清水達也, 小林直樹

    日本再生医療学会総会(Web)   18th  2019

    J-GLOBAL

  • 立体組織構築のためのバイオプリンタを用いたハイドロゲルのマイクロ加工

    田中龍一郎, 坂口勝久, 清水達也, 梅津信二郎

    日本材料科学会学術講演大会講演予稿集   2019  2019

    J-GLOBAL

  • Low-temperature culturing improves survival rate of tissue-engineered cardiac cell sheets.

    Katsuhisa Sakaguchi, Yuto Hinata, Yuki Kagawa, Kiyotaka Iwasaki, Satoshi Tsuneda, Tatsuya Shimizu, Mitsuo Umezu

    Biochemistry and biophysics reports   14   89 - 97  2018.07  [International journal]

     View Summary

    Assembling three-dimensional (3D) tissues from single cells necessitates the use of various advanced technological methods because higher-density tissues require numerous complex capillary structures to supply sufficient oxygen and nutrients. Accordingly, creating healthy culture conditions to support 3D cardiac tissues requires an appropriate balance between the supplied nutrients and cell metabolism. The objective of this study was to develop a simple and efficient method for low-temperature cultivation (< 37 °C) that decreases cell metabolism for facilitating the buildup of 3D cardiac tissues. We created 3D cardiac tissues using cell sheet technology and analyzed the viability of the cardiac cells in low-temperature environments. To determine a method that would allow thicker 3D tissues to survive, we investigated the cardiac tissue viability under low-temperature culture processes at 20-33.5 °C and compared it with the viability under the standard culture process at 37 °C. Our results indicated that the standard culture process at 37 °C was unable to support higher-density myocardial tissue; however, low-temperature culture conditions maintained dense myocardial tissue and prevascularization. To investigate the efficiency of transplantation, layered cell sheets produced by the low-temperature culture process were also transplanted under the skin of nude rats. Cardiac tissue cultured at 30 °C developed denser prevascular networks than the tissue cultured at the standard temperature. Our novel findings indicate that the low-temperature process is effective for fabricating 3D tissues from high-functioning cells such as heart cells. This method should make major contributions to future clinical applications and to the field of organ engineering.

    DOI PubMed

  • Simple action potential measurement of cardiac cell sheet utilizing electronic sheet

    Takashi Ohya, Kazuki Nakazono, Tetsutaro Kikuchi, Daisuke Sasaki, Katsuhisa Sakaguchi, Tatsuya Shimizu, Kenjiro Fukuda, Takao Someya, Shinjiro Umezu

    Artificial Life and Robotics   23   1 - 7  2018.03

     View Summary

    In this study, a device for measuring the action potential of cardiac cell sheets was developed. The action potential was measured using a device comprising a 2-µm-thick parylene film with a silver electrode printed on it, which was referred to as the “electronic sheet.” The thin parylene film exhibits high biocompatibility and flexibility. Therefore, it demonstrates promise for biomedical microelectromechanical system applications. In this study, a cell sheet was used because the interest it had garnered in regenerative medicine for creating cardiac tissue in vitro similar to that in vivo. A high-efficiency drug development system can be realized by combining cell sheet technology and fabricating a flexible electronic sheet. The action potential of a cardiac cell sheet from a rat was measured using the as-developed flexible electronic sheet.

    DOI

  • ジェランガムゲルを用いたバイオデバイス開発のためのゲル濃度検討

    田中龍一郎, 坂口勝久, 坂口勝久, 清水達也, 梅津信二郎

    日本材料科学会学術講演大会講演予稿集   2018  2018

    J-GLOBAL

  • 異なる径の管腔構造を有する生体組織の作製

    秋元渓, 坂口勝久, 坂口勝久, 清水達也, 梅津信二郎

    日本材料科学会学術講演大会講演予稿集   2018  2018

    J-GLOBAL

  • 立体臓器創製に向けた細胞シート工学

    坂口勝久, 戸部友輔, 清水達也, 梅津光生

    日本材料科学会学術講演大会講演予稿集   2018  2018

    J-GLOBAL

  • 細胞シートを用いたin vitro肝臓病態モデルの構築

    高博韜, 坂口勝久, 清水達也

    高分子学会医用高分子シンポジウム講演要旨集   47th  2018

    J-GLOBAL

  • Fabrication of gellan gum gels with unique shapes utilizing electrostatic inkjet.

    田中龍一郎, 坂口勝久, 清水達也, 梅津信二郎

    日本機械学会関東支部・精密工学会・茨城大学工学部茨城講演会講演論文集(CD-ROM)   26th  2018

    J-GLOBAL

  • IVT-mRNAを利用した目的の成長因子を分泌する細胞シート作製の検討

    秋山義勝, KWON Hyo Kyoung, 原口裕次, 坂口勝久, 大和雅之, LEE Hyukjin, 清水達也

    日本バイオマテリアル学会大会予稿集(Web)   40th  2018

    J-GLOBAL

  • 3次元浮遊攪拌によるiPS細胞の未分化増幅からのシームレス心筋分化誘導と後処理工程開発

    松浦勝久, 和田昌憲, 坂口勝久, 清水達也

    日本再生医療学会総会(Web)   17th  2018

    J-GLOBAL

  • 立体心筋組織構築のための脱細胞化ブタ小腸を用いたヒト血管床再構築法の検討

    戸部友輔, 坂口勝久, 佐野和紀, 関根秀一, 清水達也, 小林英司, 梅津光生

    日本再生医療学会総会(Web)   17th  2018

    J-GLOBAL

  • フィブリンゲルを用いた細胞シート高速積層技術による立体心筋組織の構築

    中園一紀, 坂口勝久, 清水達也, 梅津光生

    日本再生医療学会総会(Web)   17th  2018

    J-GLOBAL

  • 細胞シート工学とマイクロチタン線を用いた灌流可能な血管様構造を有する立体組織の作製

    秋元渓, 坂口勝久, 清水達也, 梅津信二郎

    日本再生医療学会総会(Web)   17th  2018

    J-GLOBAL

  • 量産した細胞凝集塊の非用手的分散を可能にするデバイス開発

    松浦勝久, 和田昌憲, 坂口勝久, 清水達也

    日本再生医療学会総会(Web)   17th  2018

    J-GLOBAL

  • 生体外で積層したヒトiPS細胞由来心筋細胞シートの収縮力とシート断面積の関係性の検討

    高橋啓明, 坂口勝久, 佐々木大輔, 松浦勝久, 清水達也, 梅津光生

    日本再生医療学会総会(Web)   17th  2018

    J-GLOBAL

  • 灌流培養床を用いた3次元組織の構築

    坂口勝久, 清水達也, 梅津光生

    日本生体医工学会大会プログラム・抄録集(Web)   57th  2018

    J-GLOBAL

  • 脱細胞化ブタ小腸を骨格としたヒトに移植可能な血管床の開発

    戸部友輔, 坂口勝久, 佐野和紀, 関根秀一, 清水達也, 小林英司, 梅津光生

    日本生体医工学会大会プログラム・抄録集(Web)   57th  2018

    J-GLOBAL

  • 細胞シート技術を用いた肝組織モデルの構築と機能評価

    森山千弘, 坂口勝久, 高博韜, 清水達也, 梅津光生

    日本再生医療学会総会(Web)   17th  2018

    J-GLOBAL

  • バイオプリンティングにおけるジェランガムのゲル化方法の開発

    田中龍一郎, 坂口勝久, 坂口勝久, 清水達也, 梅津信二郎

    日本再生医療学会総会(Web)   17th  2018

    J-GLOBAL

  • VEGF分泌を促し,またVEGF likeな作用も有する新規低分子COA-Cl

    塚本郁子, 高田麻紀, 五十嵐淳介, 榊原紀和, 窪田泰夫, 中園一紀, 坂口勝久, 清水達也, 岡野光夫, 小西良士

    日本再生医療学会総会(Web)   17th  2018

    J-GLOBAL

  • 藻類共培養による新規細胞培養法の基礎検討

    上本詩織, 梅津信二郎, 原口裕次, 坂口勝久, 清水達也

    日本機械学会関東支部総会・講演会講演論文集(CD-ROM)   24th  2018

    J-GLOBAL

  • 微小流路付きコラーゲンゲル培養床をもちいた立体組織構築法の開発

    坂口勝久, 清水達也, 梅津光生

    日本機械学会年次大会講演論文集(CD-ROM)   2018  2018

    J-GLOBAL

  • Rapid Fabrication of Tubular Tissue using Centrifuge

    上本詩織, 原口裕次, 坂口勝久, 梅津信二郎, 清水達也

    日本機械学会バイオフロンティア講演会講演論文集(CD-ROM)   29th  2018

    J-GLOBAL

  • Fabrication of 3-dimentional staked tissue for long-term hepatocytes culture

    森山千弘, 坂口勝久, GAO Botao, 清水達也, 梅津光生

    日本機械学会バイオフロンティア講演会講演論文集(CD-ROM)   29th  2018

    J-GLOBAL

  • Vascularized Tissue-Engineered Model for Studying Drug Resistance in Neuroblastoma

    A. Villasante, K. Sakaguchi, J. Kim, N.K. Cheung, M. Nakayama, H. Parsa, T. Okano, T. Shimizu, G. Vunjak-Novakovic

    Theranostics   7 ( 17 ) 4099 - 4117  2017  [Refereed]  [International journal]

     View Summary

    Neuroblastoma is a vascularized pediatric tumor derived from neural crest stem cells that displays vasculogenic mimicry and can express a number of stemness markers, such as SOX2 and NANOG. Tumor relapse is the major cause of succumbing to this disease, and properties attributed to cancer stem-like cells (CSLC), such as drug-resistance and cell plasticity, seem to be the key mechanisms. However, the lack of controllable models that recapitulate the features of human neuroblastoma limits our understanding of the process and impedes the development of new therapies. In response to these limitations, we engineered a perfusable, vascularized in vitro model of three-dimensional human neuroblastoma to study the effects of retinoid therapy on tumor vasculature and drug-resistance. METHODS: The in vitro model of neuroblastoma was generated using cell-sheet engineering and cultured in a perfusion bioreactor. Firstly, we stacked three cell sheets containing SKNBE(2) neuroblastoma cells and HUVEC. Then, a vascular bed made of fibrin, collagen I and HUVEC cells was placed onto a collagen-gel base with 8 microchannels. After gelling, the stacked cell sheets were placed on the vascular bed and cultured in the perfusion bioreactor (perfusion rate: 0.5 mL/min) for 4 days. Neuroblastoma models were treated with 10μM isotretionin in single daily doses for 5 days. RESULTS: The bioengineered model recapitulated vasculogenic mimicry (vessel-like structure formation and tumor-derived endothelial cells-TECs), and contained CSLC expressing SOX2 and NANOG. Treatment with Isotretinoin destabilized vascular networks but failed to target vasculogenic mimicry and augmented populations of CSLCs expressing high levels of SOX2. Our results suggest that CSLCs can transdifferentiate into drug resistant CD31+-TECs, and reveal the presence of an intermediate state STEC (stem tumor-derived endothelial cell) expressing both SOX2 and CD31. CONCLUSION: Our results reveal some roles of SOX2 in drug resistance and tumor relapse, and suggest that SOX2 could be a therapeutic target in neuroblastoma.

    DOI PubMed

  • 立体組織構築のための脱細胞化小腸を用いた血管床の開発

    関根秀一, 戸部友輔, 佐野和紀, 八鍬一貴, 坂口勝久, 松浦勝久, 梅津光生, 小野稔, 小林英司, 清水達也

    日本小腸移植研究会プログラム・抄録集   29th ( 2-3 ) 286 - 287  2017

    J-GLOBAL

  • 静電インクジェット法を用いたバイオ用マイクロ3Dプリンター

    梅津信二郎, 田中龍一郎, 坂口勝久, 清水達也

    再生医療   16  2017

    J-GLOBAL

  • 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

    再生医療   16  2017

    J-GLOBAL

  • 積層したヒトiPS細胞由来心筋細胞シートの収縮力測定

    高橋啓明, 坂口勝久, 佐々木大輔, 松浦勝久, 清水達也, 梅津光生

    再生医療   16  2017

    J-GLOBAL

  • 細胞シート工学とマイクロチタン線を用いた血管様構造を有する立体組織の構築

    秋元渓, 坂口勝久, 坂口勝久, 新井隆史, 新井隆史, 清水達也, 梅津信二郎

    再生医療   16  2017

    J-GLOBAL

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

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

    再生医療   16  2017

    J-GLOBAL

  • 静電インクジェットを用いた高精度なバイオマテリアルのプリント

    田中龍一郎, 田中龍一郎, 坂口勝久, 坂口勝久, 清水達也, 梅津信二郎

    再生医療   16  2017

    J-GLOBAL

  • 細胞シート技術を用いた機能的な三次元ヒト肝組織モデルの構築

    亀井翔太, 亀井翔太, 坂口勝久, 坂口勝久, 高博韜, 清水達也, 梅津光生

    再生医療   16  2017

    J-GLOBAL

  • 生体接着剤を用いた細胞シート高速積層技術による立体組織の構築と移植評価

    中園一紀, 坂口勝久, 坂口勝久, 清水達也, 梅津光生

    再生医療   16  2017

    J-GLOBAL

  • 立体管状心筋組織構築に向けたブタ脱細胞化小腸を用いた血管床の開発

    戸部友輔, 戸部友輔, 坂口勝久, 坂口勝久, 佐野和紀, 関根秀一, 清水達也, 小林英司, 梅津光生, 岡野光夫

    再生医療   16  2017

    J-GLOBAL

  • コラーゲン血管床を用いた立体心筋組織の構築

    坂口勝久, 清水達也, 松浦勝久, 大和雅之, 梅津光生, 岡野光夫

    再生医療   16  2017

    J-GLOBAL

  • 積層したヒトiPS細胞由来心筋細胞シートの厚さと収縮力の関係性の解明

    高橋啓明, 高橋啓明, 坂口勝久, 坂口勝久, 佐々木大輔, 松浦勝久, 清水達也, 梅津光生

    人工臓器(日本人工臓器学会)   46 ( 2 ) S - 120  2017

    J-GLOBAL

  • 細胞シート技術を用いた立体心筋組織構築のための脱細胞化ブタ小腸血管床の作製

    戸部友輔, 戸部友輔, 坂口勝久, 坂口勝久, 佐野和紀, 関根秀一, 清水達也, 小林英司, 梅津光生

    人工臓器(日本人工臓器学会)   46 ( 2 ) S - 121  2017

    J-GLOBAL

  • 細胞シート技術を用いたヒト立体心筋組織構築

    坂口勝久, 戸部友輔, 中園一紀, 清水達也, 梅津光生

    日本生体医工学会大会プログラム・抄録集(Web)   56th  2017

    J-GLOBAL

  • マイクロバイオ3Dプリンタを用いたバイオマテリアルゲルパターニング技術の開発

    田中龍一郎, 坂口勝久, 清水達也, 梅津信二郎

    日本生体医工学会大会プログラム・抄録集(Web)   56th  2017

    J-GLOBAL

  • マイクロバイオ3Dプリンタによるバイオマテリアルゲルのパターニング

    田中龍一郎, 坂口勝久, 清水達也, 梅津信二郎

    日本機械学会年次大会講演論文集(CD-ROM)   2017  2017

    J-GLOBAL

  • 積層したヒトiPS細胞由来心筋シートの収縮力とシート断面積の関係性の解明

    高橋啓明, 坂口勝久, 戸部友輔, 中園一紀, 佐々木大輔, 松浦勝久, 清水達也, 梅津光生

    バイオエンジニアリング講演会講演論文集(CD-ROM)   30th  2017

    J-GLOBAL

  • In vitroにおける立体組織構築を目的とした脱細胞化血管床による積層化細胞シート内血管への培養液の灌流誘導技術の検討

    戸部友輔, 坂口勝久, 高橋啓明, 中園一紀, 佐野和紀, 関根秀一, 清水達也, 小林英司, 梅津光生

    バイオエンジニアリング講演会講演論文集(CD-ROM)   30th  2017

    J-GLOBAL

  • 生体接着剤を用いた細胞シート高速積層技術による立体心筋組織の構築

    中園一紀, 坂口勝久, 高橋啓明, 戸部友輔, 中園一紀, 坂口勝久, 高橋啓明, 戸部友輔, 清水達也, 梅津光生

    バイオエンジニアリング講演会講演論文集(CD-ROM)   30th  2017

    J-GLOBAL

  • 細胞シート工学とマイクロチタン線を用いた血管構造を有する立体組織の作製

    秋元渓, 坂口勝久, 清水達也, 梅津信二郎

    日本機械学会関東支部総会・講演会講演論文集(CD-ROM)   23rd  2017

    J-GLOBAL

  • マイクロバイオ3Dプリンタの開発とバイオマテリアルのパターニング

    田中龍一郎, 坂口勝久, 清水達也, 梅津信二郎

    日本機械学会情報・知能・精密機器部門講演会論文集(CD-ROM)   2017  2017

    J-GLOBAL

  • Developing high precision patterning technology of biomaterial gels

    田中 龍一郎, 坂口 勝久, 清水 達也, 梅津 信二郎

    Imaging Conference Japan論文集   2017   13 - 16  2017

    CiNii J-GLOBAL

  • 臨床応用を目的とした立体心筋組織構築に向けた脱細胞化血管床の開発

    戸部 友輔, 坂口 勝久, 佐野 和紀, 関根 秀一, 清水 達也, 小林 英司, 梅津 光生, 岡野 光夫

    日本バイオレオロジー学会誌(B&R)   30 ( 2 ) 62 - 62  2016.06

  • 細胞シート立体積層化のための脱細胞化血管床の開発に向けた脱細胞化条件の検討

    戸部友輔, 坂口勝久, 佐野和紀, 関根秀一, 清水達也, 梅津光生, 岡野光夫

    再生医療   15  2016

    J-GLOBAL

  • フィブリンゲルとスタンプ型デバイスを用いた細胞シートの積層による3次元組織構築の加速

    日向裕人, 坂口勝久, 清水達也, 常田聡

    再生医療   15  2016

    J-GLOBAL

  • 灌流培養システムによる生体外での微小循環付きヒト三次元組織モデルの構築

    佐藤祐樹, 坂口勝久, 佐藤祐樹, 坂口勝久, 清水達也, 梅津光生, 梅津光生, 岡野光夫

    再生医療   15  2016

    J-GLOBAL

  • 細胞シートを用いた機能的な肝組織モデルおよび薬効試験デバイスの構築

    亀井翔太, 亀井翔太, 坂口勝久, 坂口勝久, 清水達也, 梅津光生, 岡野光夫

    再生医療   15  2016

    J-GLOBAL

  • マイクロ3Dプリンタを用いた着弾面の材料による吐出精度の変化

    田中龍一郎, 坂口勝久, 坂口勝久, 清水達也, 清水達也, 梅津信二郎

    Meeting of Federation of Imaging Societies (CD-ROM)   3rd  2016

    J-GLOBAL

  • 細胞シート技術を用いたヒト立体心筋組織構築のための灌流可能な血管網導入デバイスの開発

    坂口勝久, 清水達也, 松浦勝久, 大和雅之, 梅津光生, 岡野光夫

    日本生体医工学会大会プログラム・論文集(CD-ROM)   55th  2016

    J-GLOBAL

  • 臨床応用を目的とした立体心筋組織構築に向けた脱細胞化血管床の開発

    戸部友輔, 戸部友輔, 坂口勝久, 坂口勝久, 佐野和紀, 関根秀一, 清水達也, 小林英司, 梅津光生, 岡野光夫

    日本バイオレオロジー学会誌(Web)   30 ( 2 )  2016

    J-GLOBAL

  • バイオマテリアルをコートしたチタン細線における細胞接着・成長特性

    秋元渓, 坂口勝久, 清水達也, 梅津信二郎

    日本機械学会機械材料・材料加工技術講演会論文集(CD-ROM)   24th  2016

    J-GLOBAL

  • インク保温機能を搭載したマイクロ3Dプリンタによるゼラチン水溶液の吐出特性

    田中龍一郎, 坂口勝久, 清水達也, 梅津信二郎

    日本機械学会機械材料・材料加工技術講演会論文集(CD-ROM)   24th  2016

    J-GLOBAL

  • マイクロアルギン酸ゲルチューブ作製時のゼラチン溶解特性

    新井隆史, 坂口勝久, 清水達也, 梅津信二郎

    日本機械学会機械材料・材料加工技術講演会論文集(CD-ROM)   24th  2016

    J-GLOBAL

  • 3D Printing of Gelatin utilizing Micro 3D Printer

    田中 龍一郎, 坂口 勝久, 梅津 信二郎

    Imaging Conference Japan論文集   2016   7 - 9  2016

    CiNii J-GLOBAL

  • H-3-2 Fabrication of micro gelatin fibers utilizing a PTFE mold

    TANAKA Ryu-ichiro, UEHARA Yoshihiro, SAKAGUCHI Katsuhisa, UMEZU Shinjiro

    Conference on Information, Intelligence and Precision Equipment : IIP   2016   "H - 3-2-1"-"H-3-2-2"  2016

     View Summary

    Gelatin is useful biomaterials for biofabrication. The property of gelatin is unique. The state is changed to sol or gel by temperature. Utilizing the property of gelatin, we are able to fabricate cave for artificial Vessels in biodevices. Therefore, micro gelatin fibers are useful devices for fabrication of artifical vessles. In this paper, we made a mold for fabrication of micro gelatin fibers. We used PTFE for molds, because it has non-adhesive. Then, we made micro gelatin fibers which were 20〜100 μm in width utilizing the mold. Machining marks of the mold were transcribed on the surface of micro gelatin fibers. We are able to fabricate cave with arbitrary shape for artificial vessels utilizing micro gelatin fibers.

    CiNii J-GLOBAL

  • Construction of 3D Tissue by Cell Sheet Technology

    坂口勝久

    日本画像学会誌   55 ( 1 ) 83 - 87  2016

     View Summary

    Construction of three-dimensional (3D) tissues with pre-isolated cells is a promising achievement for novel medicine and drug-discovery research. Our laboratory constructs 3D tissues with an innovative and unique method for layering multiple cell sheets. Native animal tissues and organs have an abundance of capillaries to supply oxygen and nutrients, and to remove waste molecules. To construct even thicker 3D tissues by layering multiple cell sheets, it is critical to have medium or blood flow within the vascular networks of the cell sheets. Therefore, to perfuse medium in the cell sheet vascular network to maintain the viability of all cells, we developed two types of vascular beds. Such bioreactor systems, when combined with cell sheet engineering techniques, have produced functional vascularized 3D tissues. Here we explain and discuss the various processes to obtain vascular networks by properly connecting cell sheets and the engineering of 3D tissues.

    CiNii J-GLOBAL

  • Construction of Functional Liver Tissue Model by Cell Sheet Technology for Drug Test

    S. Kamei, K. Sakaguchi, L. Li, T. Shimizu, M. Umezu

    TISSUE ENGINEERING PART A   21   S264 - S264  2015.09

    Research paper, summary (international conference)  

  • Engineering Vascularized Cardiac Tissue In Vitro

    K. Sakaguchi, T. Shimizu, K. Matsuura, M. Umezu

    TISSUE ENGINEERING PART A   21   S139 - S139  2015.09

    Research paper, summary (international conference)  

  • Bioreactor technologies for myocardial tissue engineering

    Katsuhisa Sakaguchi

    Myocardial Tissue Engineering     211 - 220  2015.01

    DOI

  • Induced Pluripotent Stem Cell Differentiation under Constant Shear Stress

    N. K. Mohd Zin, K. Sakaguchi, Y. Haraguchi, T. Yagi, K. Masuura, T. Shimizu, M. Umezu

    7TH WACBE WORLD CONGRESS ON BIOENGINEERING 2015   52   7 - 10  2015

     View Summary

    Mechanotransduction in in-vitro studies has yet to be done extensively using human induced pluripotent (hiPS) cells. These external mechanical factors are important controlling factor for cell differentiation at a cellular level particularly in cardiomyocytes cells. Cells in a living body are being exposed to many kind of stresses; shear stress in particular modulates cellular function in a living body. Similarly, we are trying to understand the link between the shear stress in the bioreactor and the differentiation effects it has on iPS cells. In this study, we investigate the difference of strictly uniform laminar shear stress with a non-uniform laminar flow and its effects on the cell survival rate and differentiation. Additionally, the elimination of biochemical factors were done to satisfy the need of demand for clinically usable cells for the near future use. By eliminating cytokine induced differentiation. In addition, information pertaining to relationship between fluid shear stress, cellular deformations, cell differentiation and cell survival rate could provide a more optimised condition for cultivation specific type of cells within a shorter time period.

    DOI

  • 薬効試験に向けた機能的な肝細胞シートの作製

    亀井翔太, 亀井翔太, 李力, 李力, 坂口勝久, 坂口勝久, 清水達也, 梅津光生, 岡野光夫

    再生医療   14  2015

    J-GLOBAL

  • 低温環境下灌流培養による三次元心筋組織の構築

    松浦加奈, 松浦加奈, 日向裕人, 日向裕人, 坂口勝久, 坂口勝久, 清水達也, 梅津光生, 岡野光夫

    再生医療   14  2015

    J-GLOBAL

  • 三次元組織構築に向けた細胞シートへの血管網導入技術の開発

    坂口勝久, 清水達也, 松浦勝久, 大和雅之, 梅津光生, 岡野光夫

    再生医療   14  2015

    J-GLOBAL

  • 3次元心筋組織構築に向けた低温培養の影響評価

    日向裕人, 坂口勝久, 加川友己, 清水達也, 常田聡

    再生医療   14  2015

    J-GLOBAL

  • 細胞シートを用いた機能的な肝組織モデルおよび薬効試験デバイスの構築

    亀井翔太, 李力, 坂口勝久, 亀井翔太, 李力, 坂口勝久, 清水達也, 梅津光生, 岡野光夫

    日本バイオレオロジー学会年会プログラム・抄録集   38th ( 2 ) 69 - 69  2015

    J-GLOBAL

  • 3次元心筋組織構築に向けた低温培養の影響評価

    日向裕人, 坂口勝久, 清水達也, 常田聡

    日本バイオマテリアル学会大会予稿集   37th   129 - 129  2015

    J-GLOBAL

  • 「3Dプリンタと医療」3D組織構築のためのバイオリアクタの開発

    坂口勝久, 清水達也, 坂口勝久, 清水達也

    人工臓器(日本人工臓器学会)   44 ( 1 ) 57 - 61  2015

    CiNii J-GLOBAL

  • C212 Construction of Human Tissue Model with Microcirculation by Perfusion Culture in vitro

    SATO Yuki, SAKAGUCHI Katsuhisa, SHIMIZU Tatsuya, UMEZU Mitsuo, OKANO Teruo

    The Proceedings of the JSME Conference on Frontiers in Bioengineering   26th ( 0 ) 153 - 154  2015

    CiNii J-GLOBAL

  • 3次元組織構築のための人工血管床デバイス

    竹原宏明, 坂口勝久, 関根秀一, 清水達也, 大和雅之, 梅津光生, 岡野光夫

    応用物理学会春季学術講演会講演予稿集(CD-ROM)   61st  2014

    J-GLOBAL

  • 生体外におけるヒトiPS細胞由来心筋細胞を用いた組織構築

    田中裕也, 田中裕也, 坂口勝久, 坂口勝久, 清水達也, 関根秀一, 松浦勝久, 大和雅之, 梅津光生, 岡野光夫

    再生医療   13  2014

    J-GLOBAL

  • 3次元組織構築のためのシリコーン樹脂製人工血管床デバイスの開発

    竹原宏明, 坂口勝久, 関根秀一, 清水達也, 大和雅之, 梅津光生, 岡野光夫

    再生医療   13  2014

    J-GLOBAL

  • ヒトiPS細胞由来心筋細胞シートを用いたチューブ状立体組織の作製と環流培養装置の検討

    鶴山晋平, 坂口勝久, 清水達也, 関根秀一, 松浦勝久, 岡野光夫

    再生医療   13  2014

    J-GLOBAL

  • 低温培養および培養液灌流可能な血管網導入技術による3次元心筋組織の構築

    坂口勝久, 清水達也, 松浦加奈, 日向祐人, 関根秀一, 大和雅之, 梅津光生, 岡野光夫

    再生医療   13  2014

    J-GLOBAL

  • 循環器系DRYラボセンターを駆使した治療リスクの低減システム構築と人材育成 左冠動脈主幹部分岐狭窄血管モデルを用いたステントの血管壁への不完全圧着領域評価方法の開発

    岩崎清隆, 坂口勝久, 山崎健二, 笠貫宏

    循環器系DRYラボセンターを駆使した治療リスクの低減システム構築と人材育成に関する研究 平成25年度 総括・分担研究報告書     3  2014

    J-GLOBAL

  • 移植へ向けた動静脈閉鎖系三次元ヒト組織の構築

    佐藤祐樹, 坂口勝久, 田中裕也, 清水達也, 岩崎清隆, 大和雅之, 梅津光生, 岡野光夫

    再生医療   13   305  2014

    J-GLOBAL

  • ヒトIPS細胞由来心筋細胞シートへの灌流血管網導入技術の開発

    坂口勝久, 清水達也, 岩崎清隆, 大和雅之, 梅津光生, 岡野光夫

    日本バイオレオロジー学会年会プログラム・抄録集   37th ( 2 ) 74 - 86  2014

    J-GLOBAL

  • 2S-Ep05 Fabrication of vascularized tissue by cell sheet engineering

    Sakaguchi Katsuhisa, Shimizu Tatsuya, Yamato Masayuki, Umezu Mitsuo, Okano Teruo

    日本生物工学会大会講演要旨集   66th   106 - 106  2014

    CiNii J-GLOBAL

  • 1E43 In vitro construction of three dimensional tissue with circulation system

    SATO Yuki, SAKAGUCHI Katsuhisa, TANAKA Yuya, SHIMIZU Tatsuya, IWASAKI Kiyotaka, YAMATO Masayuki, UMEZU Mitsuo, OKANO Teruo

    The Proceedings of the Bioengineering Conference Annual Meeting of BED/JSME   2014 ( 0 ) 159 - 160  2014

    CiNii

  • Fabrication of Three-dimension Tissue by Cell Sheet Technology

    坂口 勝久, 清水 達也

    Bio industry   31 ( 1 ) 13 - 19  2014

    CiNii J-GLOBAL

  • 循環器系DRYラボセンターを駆使した治療リスクの低減システム構築と人材育成 左冠動脈主幹部分岐狭窄血管モデルを用いたステント血管壁への不完全圧着領域評価方法の開発

    岩崎清隆, 坂口勝久, 山崎健二, 笠貫宏

    循環器系DRYラボセンターを駆使した治療リスクの低減システム構築と人材育成に関する研究 平成24年度 総括・分担研究報告書    2013

    J-GLOBAL

  • 重層化細胞シートへの培養液灌流可能な血管網導入技術の開発

    坂口勝久, 清水達也, 関根秀一, 大和雅之, 梅津光生, 岡野光夫

    再生医療   12  2013

    J-GLOBAL

  • 灌流培養バイオリアクタを用いた血管網付きヒト心筋組織の構築

    中山悠衣, 清水達也, 坂口勝久, 加川友己, 松浦勝久, 常田聡, 梅津光生, 岡野光夫

    再生医療   12  2013

    J-GLOBAL

  • 立体臓器構築を目指した心筋細胞シートによるチューブ状心筋組織の作製

    鶴山晋平, 鶴山晋平, 坂口勝久, 清水達也, 関根秀一, 青木信奈子, 松浦勝久, 梅津光生, 岡野光夫

    再生医療   12  2013

    J-GLOBAL

  • 組織移植へ向けた動静脈閉鎖系心筋組織の開発

    田中裕也, 坂口勝久, 清水達也, 関根秀一, 岩崎清隆, 大和雅之, 梅津光生, 岡野光夫

    再生医療   12   260  2013

    J-GLOBAL

  • Vascularized tissue fabrication from human cells using in vitro bioreactor

    Y. Nakayama, Y. H. Lo, K. Sakaguchi, Y. Kagawa, T. Shimizu, M. Yamato, S. Tsuneda, M. Umezu, T. Okano

    JOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE   6   300 - 300  2012.09

    Research paper, summary (international conference)  

  • Engineering vascularized cardiac tissue in vitro

    K. Sakaguchi, Y. Tanaka, H. Sekine, T. Shimizu, M. Yamato, M. Umezu, T. Okano

    JOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE   6   353 - 353  2012.09

    Research paper, summary (international conference)  

  • Fabrication of three-dimensional tubular cardiac tissue using neonatal mouse cardiomyocytes

    S. Tsuruyama, K. Sakaguchi, T. Shimizu, H. Sekine, S. Aoki, K. Matsuura, M. Umezu, T. Okano

    JOURNAL OF TISSUE ENGINEERING AND REGENERATIVE MEDICINE   6   354 - 354  2012.09

    Research paper, summary (international conference)  

  • 細胞シート積層による三次元心筋組織構築過程における代謝評価

    加川 友己, 田中 裕也, 坂口 勝久, 関根 秀一, 清水 達也, 常田 聡, 岡野 光夫

    組織培養研究   31 ( 1 ) 41 - 41  2012.03

  • Novel device for transplantation of cell sheet and evaluation of thin polymer films by atomic force microscopy

    Ryohei Takeuchi, Kazuhiro Fukumori, Katsuhisa Sakaguchi, Yutaka Terajima, Tatsuya Shimizu, Teruo Okano, Mitsuo Umezu

    2011 Int. Symp. on Micro-NanoMechatronics and Human Science, Symp. on "COE for Education and Research of Micro-Nano Mechatronics", Symposium on "Hyper Bio Assembler for 3D Cellular System Innovation"     168 - 173  2012

     View Summary

    Cell therapy is expected to a new tool to treat refractory diseases. In heart regeneration, it has been firstly conducted with needle injection of cell suspensions. Recently, cell sheet engineering emerged as another method of cell therapy. Cell sheet is prepared with a temperature responsive dish by temperature reduction. It is a thin-patch-like tissue construct and its thickness is several tens of micrometers. It is composed of cells and intrinsic extra cellular matrix only. The transplantation of the cell sheet has been already conducted in animal experiments and even in clinical trials. The cell sheet is transplanted at the surface of the heart, but it is difficult to transplant the cell sheet under the beating heart. To overcome this difficulty, we designed a device that was composed of two thin polymer films that have different friction. The films were made of polyurethane, polyethylene, or polypropylene. The cell sheet was set up on the device by sandwiching it with the less frictional film and the more frictional film. In this paper, using two different films having the different friction, the cell sheet was successfully transplanted to the static round polymer surface, the harvested heart, and even the beating heart of pig by removing the films step by step using the difference in friction. Also, surface properties such as friction, adhesion force and roughness of the films were studied by an atomic force microscopy (AFM). From the results of the study, the friction of the film was found to be likely proportional to the adhesion force and the inverse of roughness. © 2011 IEEE.

    DOI

  • 培地・酸素供給の変化に応じた細胞シートの代謝変動特性

    加川友己, 前山絵里奈, 関根和希, 坂口勝久, 関根秀一, 清水達也, 常田聡, 岡野光夫

    再生医療   11  2012

    J-GLOBAL

  • マウス胎児由来心筋細胞シートを用いたチューブ状三次元組織の構築

    鶴山晋平, 鶴山晋平, 坂口勝久, 清水達也, 関根秀一, 青木信奈子, 松浦勝久, 梅津光生, 岡野光夫

    再生医療   11  2012

    J-GLOBAL

  • 多層化心筋細胞シートへの血管導入技術の検討

    坂口勝久, 清水達也, 関根秀一, 大和雅之, 梅津光生, 岡野光夫

    再生医療   11  2012

    J-GLOBAL

  • 心筋細胞シートにおけるエネルギー代謝

    加川友己, 坂口勝久, 関根秀一, 清水達也, 常田聡, 岡野光夫

    酸素ダイナミクス研究会プログラム・抄録集   16th  2012

    J-GLOBAL

  • 心筋細胞シートによるチューブ状組織モデルの構築と内圧負荷のための灌流培養装置の開発

    鶴山晋平, 鶴山晋平, 坂口勝久, 清水達也, 関根秀一, 青木信奈子, 松浦勝久, 梅津光生, 岡野光夫

    人工臓器(日本人工臓器学会)   41 ( 2 ) S - 193  2012

    J-GLOBAL

  • 単層細胞シートの代謝評価と酸素供給の影響

    加川友己, 前山絵里奈, 関根和希, 坂口勝久, 関根秀一, 清水達也, 常田聡, 岡野光夫

    化学工学会年会研究発表講演要旨集(CD-ROM)   77th  2012

    J-GLOBAL

  • 生体外における血管網付ヒト三次元組織の構築

    中山悠衣, 洞口重人, 坂口勝久, 加川友己, 清水達也, 大和雅之, 常田聡, 梅津光生, 岡野光夫

    再生医療   11  2012

    J-GLOBAL

  • 循環器系DRYラボセンターを駆使した治療リスクの低減システム構築と人材育成 治療機器の実臨床に即した非臨床評価による適正使用法の確立 補助人工心臓装着患者の管理基準の確保に向けた循環シミュレータの確立

    岩崎清隆, 坂口勝久, 山崎健二, 笠貫宏

    循環器系DRYラボセンターを駆使した治療リスクの低減システム構築と人材育成に関する研究 平成23年度 総括・分担研究報告書     3  2012

    J-GLOBAL

  • 生体外における動静脈閉鎖系心筋組織構築

    田中裕也, 清水達也, 関根秀一, 坂口勝久, 洞口重人, 岩崎清隆, 大和雅之, 梅津光生, 岡野光夫

    再生医療   11   250  2012

    J-GLOBAL

  • 厚い心筋組織構築に向けた血管導入技術の検討

    坂口勝久, 清水達也, 岩崎清隆, 大和雅之, 梅津光生, 岡野光夫

    人工臓器(日本人工臓器学会)   41 ( 2 ) S.220 - 220  2012

    J-GLOBAL

  • 移植可能な組織片開発へ向けた動静脈閉鎖系心筋組織構築

    田中裕也, 坂口勝久, 清水達也, 関根秀一, 岩崎清隆, 大和雅之, 梅津光生, 岡野光夫

    人工臓器(日本人工臓器学会)   41 ( 2 ) S.113 - 113  2012

    J-GLOBAL

  • 7F44 The creation of a novel bioreactor for constructing microvascularized 3D tissue

    HORAGUCHI Shigeto, SHIMIZU Tatsuya, SAKAGUCHI Katsuhisa, NAKAYAMA Yui, IWASAKI Kiyotaka, YAMATO Masayuki, OKANO Teruo, UMEZU Mitsuo

    The Proceedings of the Bioengineering Conference Annual Meeting of BED/JSME   2012 ( 0 ) _7F44 - 1_-_7F44-2_  2012

    CiNii

  • 微小流路付きコラーゲンゲル培養における血管網導入三次元心筋組織の構築

    坂口勝久, 田中裕也, 洞口重人, 清水達也, 岩崎清隆, 大和雅之, 梅津光生, 岡野光夫

    日本バイオマテリアル学会大会予稿集   33rd   187 - 187  2011.11

    J-GLOBAL

  • コラーゲンゲル培養床を用いた血管網付ヒト三次元組織の基礎検討

    中山 悠衣, 洞口 重人, 坂口 勝久, 加川 友己, 清水 達也, 大和 雅之, 常田 聡, 梅津 光生, 岡野 光夫

    日本バイオマテリアル学会大会予稿集   33回   96 - 96  2011.11

  • 三次元心筋組織構築に向けたバイオリアクターの創出

    洞口重人, 洞口重人, 清水達也, 関根秀一, 坂口勝久, 坂口勝久, 小倉伸也, 小倉伸也, 大和雅之, 梅津光生, 岡野光夫

    再生医療   10  2011

    J-GLOBAL

  • In vitroにおける血管網付ヒト3次元組織の構築

    小倉伸也, 小倉伸也, 坂口勝久, 坂口勝久, 関根秀一, 清水達也, 大和雅之, 梅津光生, 岡野光夫

    再生医療   10  2011

    J-GLOBAL

  • 機械的刺激による細胞シートの挙動変化

    植木光樹, 植木光樹, 植木光樹, 坂口勝久, 清水達也, 大和雅之, 梅津光生, 梅津光生, 岡野光夫

    人工臓器(日本人工臓器学会)   40 ( 2 )  2011

    J-GLOBAL

  • 微小血管流路デバイスを用いた生体内組織構築の検討

    植木光樹, 坂口勝久, 洞口重人, 小倉伸也, 清水達也, 大和雅之, 梅津光生, 梅津光生, 岡野光夫

    再生医療   10  2011

    J-GLOBAL

  • 灌流培養における3次元心筋組織の構築

    坂口勝久, 清水達也, 岩崎清隆, 大和雅之, 梅津光生, 岡野光夫

    日本バイオレオロジー学会年会プログラム・抄録集   34th   85  2011

    J-GLOBAL

  • S021022 Fabrication of Vascularized Cardiac Tissue

    SAKAGUCHI Katsuhisa, SHIMIZU Tatsuya, IWASAKI Kiyotaka, YAMATO Masayuki, UMEZU Mitsuo, OKANO Teruo

    The Proceedings of Mechanical Engineering Congress, Japan   2011 ( 0 ) _S021022 - 1-_S021022-5  2011

     View Summary

    Recently, researchers have challenged to create three-dimensional (3-D) tissues in order to establish in vitro models and new therapy for damaged organ. We have developed cell-sheet and successfully fabricated pulsatile 3-D myocardial tissues by stacking myocardial cell sheets. However, in vitro scaling up of 3-D tissues is limited due to lack of vessels supplying oxygen and nutrition, removing waste molecules. In this study, we have developed novel bioreactor culturing cell sheets on collagen-based microchannels and examined cell behavior between tissues and channels. Rat cardiac cells including endothelial cells cultured on a temperature responsible culture dish for 4 days. By lowering temperature, confluent cardiac cells were harvested as an intact cell sheet. Collagen-based microchannels were engineered by gelling collagen around parallel stainless wires and extracting the wires. The multi-layered cell sheet was put on the collagen-gel base with microchannels and the device was connected to the novel bioreactor system. After 5 days of cultivation, rat blood was perfused into the microchannels for checking the flow and the tissue sections were stained with Hematoxylin-Eosin. HE staining demonstrated that layered cell sheets tightly connected onto the collagen microchannels. The cells migrated into collagen gel and formed like capillaries. Some cells reached to the microchannels and covered over inner surface. The perfused blood cells flew the capillaries between cell sheets and the collagen-based microchannels. We have successfully fabricated 3-D tissues with perfused vascularized capillaries. These results showed new insights regarding in vitro vascular formation and indicated the possibility of fabricating vascularized 3-D tissues.

    CiNii J-GLOBAL

  • 心筋細胞シートへの血管導入技術の開発

    坂口勝久, 清水達也, 小倉伸也, 洞口重人, 岩崎清隆, 大和雅之, 梅津光生, 岡野光夫

    日本シミュレーション学会大会発表論文集   29th   221 - 224  2010.06

    J-GLOBAL

  • “細胞から心臓を創る”~血管網導入技術の開発~

    坂口勝久, 清水達也, 岩崎清隆, 大和雅之, 梅津光生, 岡野光夫

    人工臓器(日本人工臓器学会)   39 ( 2 ) S.37  2010

    J-GLOBAL

  • 0215 Fabrication of 3-dimensional tissue using human cells

    NOGUCHI Shinsuke, OKANO Teruo, SHIMIZU Tatsuya, HORAGUCHI Shigeto, OGURA Shinya, SAKAGUCHI Katsuhisa, SEKINE Hidekazu, IWASAKI Kiyotaka, YAMATO Masayuki, UMEZU Mitsuo

    The Proceedings of the Bioengineering Conference Annual Meeting of BED/JSME   22nd ( 0 )  2010

    CiNii J-GLOBAL

  • Development of Bioreactor System for Bioengineering Three-Dimensional Myocardial Tissues

    SAKAGUCHI Katsuhisa, SAWADA Richi, SHIMIZU Tatsuya, IWASAKI Kiyotaka, YAMATO Masayuki, UMEZU Mitsuo, OKANO Teruo

    Journal of Life Support Engineering   21 ( 3 ) 104 - 109  2009.09

     View Summary

    Recently, researchers have challenged to create three-dimensional (3-D) tissues with tissue engineering technology in order to establish in vitro models and new therapy for damaged organ. Most popular approach of tissue engineering is using 3-D biodegradable scaffolds as alternatives of extracellular matrix. By contrast, we have bioengineered pulsatile myocardial tissues by stacking cardiomyocyte sheets, which were harvested from temperature-responsive culture dishes only by lowering temperature. However, the shortage of oxygen and nutrition limits the final tissue thickness. In this study, we tried to fabricate thicker myocardial tissues by promoting oxygen and nutrition permeation using a novel perfusion bioreactor. Triple-layer neonatal rat cardiomyocyte sheets were attached in the culture chamber. Chambers were perfused with 2.4mL/h culture media and 15mmHg pressure gradient was applied to perfuse the culture media through the construct. After 5-days culture, the construct in the bioreactor was thicker and cell-denser than in static condition. These results indicate that the perfusion bioreactor should contribute to myocardial tissue engineering in vitro.

    CiNii

  • Fabrication of Three-Dimensional Tissues with Perfused Microchannels

    Katsuhisa Sakaguchi, Tatsuya Shimizu, Kiyotaka Iwasaki, Masayuki Yamato, Mitsuo Umezu, Teruo Okano

    13TH INTERNATIONAL CONFERENCE ON BIOMEDICAL ENGINEERING, VOLS 1-3   23 ( 1-3 ) 1213 - +  2009

     View Summary

    Recently, researchers have challenged to create three-dimensional (3-D) tissues with tissue engineering technology in order to establish in vitro models and new therapy for damaged organ. We have developed cell-sheet based tissue engineering and successfully fabricated pulsatile 3-D myocardial tissues both in vivo and in vitro by layering cardiac cell sheets. However, in vitro scaling up of 3-D cell-dense tissues is limited due to lack of blood vessels supplying oxygen and nutrition and removing waste molecules. In this study, we have developed novel bioreactor culturing layered cell sheets on collagen-based microchannels and examined cell behavior between tissues and channels. Rat cardiac cells including endothelial cells were cultured on temperature responsible culture dishes for 4 days. By lowering temperature, confluent cardiac cells were harvested as an intact cell sheet and two cardiac cell sheets are layered. Collagen-based microchannels were engineered by gelling collagen around parallel stainless wires and extracting the wires. The double-layer cell sheets were put on the microchannels and the constructs were connected to the novel perfusion bioreactor. After 5 days of cultivation, the tissue sections were stained with Hematoxylin-Eosin and endothelial cell specific Isolectin B4. HE staining demonstrated that layered cell sheets tightly connected onto the collagen microchannels. The microchannels maintained their patency during culture period. The cardiac cells migrated into collagen gel and the number of migration increased flow-rate dependently. At higher flow-rate, some cardiac cells reached to microchannels and covered over their inner surface. Isolectin B4 staining showed endothelial cells formed networks within the cell sheets and also played as migrating cells. We have successfully fabricated 3-D tissues with perfused microchannels and tissue-originated cells migrated and communicated with the microchannels. These results showed new insights regarding in vitro vascular formation and indicated the possibility for fabricating vascularized 3-D tissues.

    DOI

  • 細胞シートが心臓に接合するまでの時間についての検討

    常徳, 清水達也, 原口裕次, 坂口勝久, 梅津光生, 岡野光夫

    再生医療   8  2009

    J-GLOBAL

  • 微小流路付きコラーゲンゲル培養床に用いた三次元心筋組織構築の検討

    坂口勝久, 清水達也, 岩崎清隆, 大和雅之, 梅津光生, 岡野光夫

    人工臓器(日本人工臓器学会)   38 ( 2 ) S.154  2009

    J-GLOBAL

  • 三次元心筋組織構築のための微小流路付きコラーゲンゲル培養の検討

    坂口勝久, 清水達也, 岩崎清隆, 大和雅之, 梅津光生, 岡野光夫

    日本バイオマテリアル学会大会予稿集   31st   373  2009

    J-GLOBAL

  • Myocardial Tissue Engineering for Damaged Heart Repair

    東京女子医科大学雑誌   79 ( 3 ) 108 - 112  2009

     View Summary

    Regeneration therapy has currently emerged as one of the most promising treatments for patients suffering from heart failure. Recently, transplantation of engineered myocardial tissues has been pursued to overcome the disadvantages of isolated cell injections. We have exploited novel cell manipulation techniques to construct 3-D cell-dense myocardial tissues by layering cell sheets, which can be harvested from temperature-responsive culture dishes by merely lowering the temperature. The stacked rat cardiomyocyte sheets simultaneously beat in macroscopic view and survived in vivo, suggested their potential to improve cardiac function. Adipose tissue-derived myoblast sheets and mesenchymal stem cell sheets as well as cardiomyocyte sheets also improved heart function. Recent developments have promoted the clinical application of autologous myoblast sheet transplantation. As the next step, we attempted to introduce blood vessels within bioengineered myocardial tissues to overcome size-limitation. A multi-step transplantation of triple-layer cell sheets was performed and finally, after the 10th transplantation, functional myocardial tissue approximately1 mm-thick, was obtained. Furthermore, we have confirmed the effectiveness of several bioreactive agents which enable media-perfusion through myocardial constructs and can stretch to imitate the in vivo environment. We believe that interdisciplinary research based on cell sheet engineering would have enormous potential in myocardial tissue regeneration.

    CiNii J-GLOBAL

  • 積層化細胞シートにおける酸素濃度解析

    坂口勝久, 清水達也, 野口慎介, 岩崎清隆, 大和雅之, 梅津光夫, 岡野光夫

    再生医療   7   234  2008

    J-GLOBAL

  • ブタ皮下脂肪組織由来間葉系幹細胞シートの作製と積層化

    常徳華, 清水達也, 原口裕次, 坂口勝久, 大和雅之, 梅津光生, 岡野光夫

    再生医療   6  2007

    J-GLOBAL

  • 細胞シートの心臓への移植手技の最適化についての検討

    CHANG Dehua., 清水達也, 原口裕次, 坂口勝久, 梅津光生, 岡野光夫

    日本心臓血圧研究振興会研究業績集   ( 22 )  2007

    J-GLOBAL

  • Development of the perfusion bioreactor to reconstruct 3-dimensional cardiac muscle tissues

    SAKAGUCHI Katsuhisa, SHIMIZU Tatsuya, IWASAKI Kiyotaka, UMEZU Mitsuo, OKANO Teruo, SAKURAI Yasuhisa

    The Proceedings of the Bioengineering Conference Annual Meeting of BED/JSME   16th ( 0 ) 445 - 446  2004

    CiNii J-GLOBAL

▼display all

Research Projects

  • 3次元組織工学による次世代食肉生産技術の創出

    国立研究開発法人 科学技術振興機構(JST)  未来社会創造事業

    Project Year :

    2020.04
    -
    2025.03
     

  • 新規不妊治療法開発のための受精卵着床子宮内膜シートの構築

    AMED  橋渡し研究 異分野融合型研究シーズ(シーズH)

    Project Year :

    2021.04
    -
    2022.03
     

    坂口勝久

  • 藻類と動物細胞を用いたサーキュラーセルカルチャーによるバイオエコノミカルな培養食料生産システム

    内閣府  ムーンショット型研究開発制度 目標 食料供給の拡大と地球環境保全を両立する食料生産システムの開発

    Project Year :

    2020.12
    -
    2022.03
     

    坂口勝久

  • Development of a minimally invasive electronic sheet for ex vivo monitoring of artificial cardiomyocyte tissue

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Challenging Research (Exploratory)

    Project Year :

    2020.07
    -
    2022.03
     

  • Real-time quantitative prediction of cerebral-aneurysm growth with thrombosis-stratified vascular remodeling

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

    Project Year :

    2019.04
    -
    2022.03
     

  • 効果的な細胞移植を実現するための細胞シート高速積層法の開発

    日本学術振興会  科学研究費助成事業 基盤研究(C)

    Project Year :

    2019.04
    -
    2022.03
     

    坂口 勝久

  • Development of Smart Electronics Sheet Within Cardiac Cell Sheet

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Challenging Research (Exploratory)

    Project Year :

    2018.06
    -
    2020.03
     

  • Controlling sheer stress in a suspension culture using qouette flow for efficient cell proliferation

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Young Scientists (B)

    Project Year :

    2017.04
    -
    2019.03
     

    Sakaguchi Katsuhisa

     View Summary

    Biopharmaceuticals manufactured by applying genetic engineering, and regenerative medicine that creates tissues and organs from stem cells and treats them are drawing attention as the next generation medical treatment. However, the next generation medical treatment will be costed enormously and will be a socially severe problem. Therefore, in this research, we developed a novel culture method to easily grow a large number of cells by performing rotational suspension culture using Couette flow. Couette flow is a flow that occurs in the liquid in the gap between two concentric rotating cylinders, and is a very stable flow that generates uniform shear stress at all locations in the incubator. As a result, by performing the developed Couette flow culture, it succeeded in cell proliferation comparable to the normal culture dish. We have found a novel culture method for growing a large number of cells using this more convenient rotary suspension culture.

  • 3D Cell Structure Utilizing Micro 3D Printer and Cell Sheet

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Challenging Exploratory Research

    Project Year :

    2016.04
    -
    2018.03
     

    Umezu Shinjiro

     View Summary

    Recently, 3D cell structures are fabricated utilizing many kinds of biofabrication technologies. Cells those are located inside of the 3D structure get nutrition via capillary blood vessels. However, since the capillary is narrow, amount of nutrition is limited. In this study we fabricated cell structures those have arterioles inside.

  • Development of a tissue with in vivo regeneration capability for reconstruction of anterior cruciate ligament using decellularization and tissue-sterilization technologies

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research (B)

    Project Year :

    2014.04
    -
    2017.03
     

    Iwasaki Kiyotaka

     View Summary

    In the treatment of knee anterior cruciate ligament (ACL) ruptured in sports and so on, patient’s own tendon is harvested and used. In this study, as a new tissue substitute, porcine tendon was used, and decellularization and sterilization technologies of tissues were established. Experiments of reconstruction of ovine knee ACL using decellularized and sterilized porcine tendons revealed that the decellularized tissues were functioned 3 months. The tissues were repopulated in vivo with ovine autologous cells and vessels were formed. Ultimate tensile loads after 3 months implantation were comparable between decellularized porcine tissues and autologous Achilles tendons. This study opened a new way to develop a novel medical device to treat ACL without need of harvesting patient’s own tendon.

  • Fabrication of structually comlicated soft tissue such as liver-tissues and understanding its biological function of teh fabricated liver-tissue

    Japan Society for the Promotion of Science  Grants-in-Aid for Scientific Research Grant-in-Aid for Scientific Research on Innovative Areas (Research in a proposed research area)

    Project Year :

    2011.04
    -
    2016.03
     

    YAMATO MASAYUKI, AKIYAMA Yoshikatsu, NAKAYAMA Masamichi, KOBAYASHI Jun, NAGASE Kenichi, TAKAHAHI Hironobu, SHIMIZU Tatsuya

     View Summary

    We have newly developed heparin-immobilized temperature-responsive cell culture surface (TRCS), further modifying the surfaces with growth factor to fabricate hepatic-cells sheet. We have demonstrated that hepatic-cells sheet was readily fabricated with growth-factor and heparin complex immobilized surface, maintaining physiological function characteristic of hepatic-tissue for longer period. Micro-patterned TRCS was also developed, being applied to fabricate nerve-tissue. The surface would be expected as base technology for fabrication of nerve-tissue. A facile method of preparation of TRCS without special equipment and reagents were newly developed. We also innovated robotics to create robot, which attained precise and multiple micro-contacting, and devise to transplant and/or carry cell-sheet easily for short time. Rapid and convenient method for fabricating soft-tissue such as hepatic and nerve tissues would be attained, combing those techniques.

  • 自己治癒能力を引き出す無細胞化組織実用化のための総合的基礎研究

    日本学術振興会  科学研究費助成事業 基盤研究(B)

    Project Year :

    2012
     
     
     

    岩崎 清隆, 坂口 勝久, 梅津 光生, 加藤 義治, 伊藤 匡史

     View Summary

    本研究は,自己治癒能力を引き出し,体内で自己細胞が入り込み自己組織化が期待できる再生促進型無細胞化腱を独自の組織無細胞化技術を駆使して開発するものである.実用化を目指した総合的基礎研究の中で,コア技術として,力学的特性の維持を実現する組織の保存・滅菌法が未解決課題であった.無細胞化組織を凍結乾燥して滅菌し,手術場で生理食塩水で再水和して使用するコンセプトで研究を推進している.初年度は,水との結合に着目して組織前処理法について詳細に検討し,凍結乾燥してエチレンオキサイドガス滅菌すると変化してしまう組織の粘弾性特性を未処理組織と同等に保持できる組織前処理条件を確立した(国際特許出願).また,我々が開発した非臨床超急性免疫反応評価法を駆使し,無細胞化処理して滅菌処理したウシ由来心膜に対するヒト血液の超急性免疫反応を,臨床で使用されており安全性が確認されているグルタールアルデヒド溶液で処理されたウシ心のう膜パッチ製品,及び,ポジティブコントロールとして未処理ウシ心膜を抗生物質で洗浄した組織と比較評価した.実験前に再水和させた無細胞化及び滅菌済みウシ由来心膜と共培養した血液のラクトフェリン濃度は,グルタールアルデヒド固定処理されたウシ心のう膜パッチと同程度であった.C3a濃度も同様の結果となり,補体活性が抑制されることを定量的に示すことができた.さらにsC5b-9濃度は,グルタールアルデヒド処理されたウシ心のう膜製品と比較して,無細胞化処理して滅菌したウシ心膜の方が活性が抑制されることが判明した.合わせて,本実験系で,未処理組織はいずれの指標も顕著に上昇することを確認した.無細胞化処理後に確立した方法で前処理し,凍結乾燥してエチレンオキサイドガス滅菌したウシ腱を用い,第一例目の大動物前十字靭帯再建術をヒツジで実施し,6ヶ月間問題なく機能することを実証できた.

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