The Experts below are selected from a list of 26142 Experts worldwide ranked by ideXlab platform
Teruo Okano - One of the best experts on this subject based on the ideXlab platform.
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Contractile force measurement of human induced pluripotent stem Cell-derived Cardiac Cell sheet-tissue.
PLOS ONE, 2018Co-Authors: Daisuke Sasaki, Katsuhisa Matsuura, Yuji Haraguchi, Hiroyoshi Seta, Teruo Okano, Tatsuya ShimizuAbstract:We have developed our original tissue engineering technology “Cell sheet engineering” utilizing temperature-responsive culture dishes. The Cells are confluently grown on a temperature-responsive culture dish and can be harvested as a Cell sheet by lowering temperature without enzymatic digestion. Cell sheets are high-Cell-density tissues similar to actual living tissues, maintaining their structure and function. Based on this “Cell sheet engineering”, we are trying to create functional Cardiac tissues from human induced pluripotent stem Cells, for regenerative therapy and in vitro drug testing. Toward this purpose, it is necessary to evaluate the contractility of engineered Cardiac Cell sheets. Therefore, in the present study, we developed a contractile force measurement system and evaluated the contractility of human iPSC-derived Cardiac Cell sheet-tissues. By attaching the Cardiac Cell sheets on fibrin gel sheets, we created dynamically beating Cardiac Cell sheet-tissues. They were mounted to the force measurement system and the contractile force was measured stably and clearly. The absolute values of contractile force were around 1 mN, and the mean force value per cross-sectional area was 3.3 mN/mm2. These values are equivalent to or larger than many previously reported values, indicating the functionality of our engineered Cardiac Cell sheets. We also confirmed that both the contractile force and beating rate were significantly increased by the administration of adrenaline, which are the physiologically relevant responses for Cardiac tissues. In conclusion, the force measurement system developed in the present study is valuable for the evaluation of engineered Cardiac Cell sheet-tissues, and for in vitro drug testing as well.
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creation of mouse embryonic stem Cell derived Cardiac Cell sheets
Biomaterials, 2011Co-Authors: Katsuhisa Matsuura, Tatsuya Shimizu, Yuji Haraguchi, Shinako Masuda, Noriko Yasuda, Nobuhisa Hagiwara, Peter W Zandstra, Teruo OkanoAbstract:Abstract Research on heart tissue engineering is an exciting and promising area. Although we previously developed bioengineered myocardium using Cell sheet-based tissue engineering technologies, the issue of appropriate Cell sources remained unresolved. In the present study, we created Cell sheets of mouse embryonic stem (ES) Cell-derived cardiomyocytes after expansion in three-dimensional stirred suspension cultures. Serial treatment of the suspension cultures with noggin and granulocyte colony-stimulating factor significantly increased the number of cardiomyocytes by more than fourfold compared with untreated cultures. After drug selection for ES Cells expressing the neomycin-resistance gene under the control of the α-myosin heavy chain promoter, almost all of the Cells showed spontaneous beating and expressed several Cardiac contractive proteins in a fine striated pattern. When ES-derived cardiomyocytes alone were seeded onto temperature-responsive culture dishes, Cell sheets were not created, whereas cocultures with Cardiac fibroblasts promoted Cell sheet formation. The cardiomyocytes in the Cell sheets beat spontaneously and synchronously, and expressed connexin 43 at the edge of adjacent cardiomyocytes. Furthermore, when the extraCellular action potential was recorded, unidirectional action potential propagation was observed. The present findings suggest that stirred suspension cultures with appropriate growth factors are capable of producing cardiomyocytes effectively and easily, and that ES-derived Cardiac Cell sheets may be a promising tool for the development of bioengineered myocardium.
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Cardiac Cell sheet transplantation improves damaged heart function via superior Cell survival in comparison with dissociated Cell injection
Tissue Engineering Part A, 2011Co-Authors: Hidekazu Sekine, Katsuhisa Matsuura, Tatsuya Shimizu, Masayuki Yamato, Nobuhisa Hagiwara, Izumi Dobashi, Masafumi Takahashi, Eiji Kobayashi, Teruo OkanoAbstract:Regenerative therapies have currently emerged as one of the most promising treatments for repair of the damaged heart. Recently, numerous researchers reported that isolated Cell injection treatments can improve heart function in myocardial infarction models. However, significant Cell loss due to primary hypoxia or Cell wash-out and difficulty to control the location of the grafted Cells remains problem. As an attempt to overcome these limitations, we have proposed Cell sheet-based tissue engineering, which involves stacking confluently cultured Cells (two-dimensional), Cell sheets, to construct three-dimensional Cell-dense tissues. Cell sheet transplantation has been able to recover damaged heart function. However, no detailed analysis for transplanted Cell survival has been previously performed. The present study compared the survival of Cardiac Cell sheet transplantation to direct Cell injection in a rat myocardial infarction model. Luciferase-expressing neonatal rat Cardiac Cells were harvested as Cell...
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endothelial Cell coculture within tissue engineered cardiomyocyte sheets enhances neovascularization and improves Cardiac function of ischemic hearts
Circulation, 2008Co-Authors: Hidekazu Sekine, Tatsuya Shimizu, Sachiko Sekiya, Masayuki Yamato, Eiji Kobayashi, Kyoko Hobo, Joseph Yang, Hiromi Kurosawa, Teruo OkanoAbstract:Background— Regenerative therapies, including myocardial tissue engineering, have been pursued as a new possibility to repair the damaged myocardium, and previously the transplantation of layered cardiomyocyte sheets has been shown to be able to improve Cardiac function after myocardial infarction. We examined the effects of promoting neovascularization by controlling the densities of cocultured endothelial Cells (ECs) within engineered myocardial tissues created using our Cell sheet-based tissue engineering approach. Methods and Results— Neonatal rat cardiomyocytes were cocultured with GFP-positive rat-derived ECs on temperature-responsive culture dishes. Cocultured ECs formed Cell networks within the cardiomyocyte sheets, which were preserved during Cell harvest from the dishes using simple temperature reduction. We also observed significantly increased in vitro production of vessel-forming cytokines by the EC-positive Cardiac Cell sheets. After layering of 3 Cardiac Cell sheets to create 3-dimensional ...
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bioengineered Cardiac Cell sheet grafts have intrinsic angiogenic potential
Biochemical and Biophysical Research Communications, 2006Co-Authors: Sachiko Sekiya, Tatsuya Shimizu, Masayuki Yamato, Akihiko Kikuchi, Teruo OkanoAbstract:Abstract Previously, we have demonstrated the long-term survival of myocardial Cell sheet constructs in vivo, with microvascular network formation throughout the engineered tissues. The understanding and control of these vascularization processes are a key factor for creating thicker functional tissues. Here, we show that Cardiac Cell sheets express angiogenesis-related genes and form endothelial Cell networks in culture. After non-invasive harvest and stacking of Cell sheets using temperature-responsive culture dishes, these endothelial Cell networks are maintained and result in neovascularization upon in vivo transplantation. Interestingly, we also discovered that all of the graft vessels are derived from the grafts themselves and these vessels migrate to connect with the host vasculature. Finally, blood vessel formation within the grafts can be controlled by changing the ratio of endothelial Cells. In conclusion, myocardial tissue grafts engineered with Cell sheet technology have their own inherent potential for the in vivo neovascularization that can be regulated in vitro.
Tatsuya Shimizu - One of the best experts on this subject based on the ideXlab platform.
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Engineering of functional Cardiac tubes by stepwise transplantation of Cardiac Cell sheets onto intestinal mesentery
Heart and Vessels, 2020Co-Authors: Noriyasu Masuda, Hidekazu Sekine, Hiroshi Niinami, Tatsuya ShimizuAbstract:Implantable organ-like grafts made using tissue engineering techniques could potentially be used as circulatory assist devices in people with heart failure. The aims of this study were to engineer implantable, thick Cardiac tubes by the stepwise transplantation of Cardiac Cell sheets onto intestinal mesentery and confirm that these Cardiac tubes exhibited pulsatile activity and generated an internal pressure. Cell sheets were created by culturing neonatal rat Cardiac Cells on temperature-responsive dishes. After harvesting, three Cell sheets were stacked, and the triple-layered Cell sheet was rolled around a section of endotracheal tube. The resulting construct was cultured to generate a Cardiac tube. In the single-step group ( n = 6), a Cardiac tube was implanted onto the intestinal mesentery of a rat. In the double-step group ( n = 6), a Cardiac tube was implanted onto the intestinal mesentery of a rat, and another new Cardiac tube was inserted into the original Cardiac tube one day later. The pulsations and internal pressures of the implanted Cardiac tubes were evaluated 1, 2 and 4 weeks after transplantation. Histology and immunohistochemistry were used to confirm whether vasculature was present in the Cardiac tubes at 4 weeks after transplantation. We found that the Cardiac tubes developed spontaneous pulsations from 1 week after transplantation. The average internal pressures of the Cardiac tubes at 4 weeks after transplantation were 1.8 ± 1.0 mmHg in the single-step group and 2.5 ± 0.3 mmHg in the double-step group. The Cardiac tubes in the double-step group contracted in response to electrical stimulation at 4 weeks after transplantation. Histological and immunohistochemical analyses revealed engraftment of the transplanted Cardiac Cell sheets and neovascularization of the Cardiac tubes in both groups. Our findings demonstrate that it is feasible to generate functional Cardiac tubes using the mesentery as a vascular bed. Further development of this technique will include the creation of a thicker tube, transplantation of the tube into major vessels and evaluation of the function of the tube under physiological conditions.
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Contractile force measurement of human induced pluripotent stem Cell-derived Cardiac Cell sheet-tissue.
PLOS ONE, 2018Co-Authors: Daisuke Sasaki, Katsuhisa Matsuura, Yuji Haraguchi, Hiroyoshi Seta, Teruo Okano, Tatsuya ShimizuAbstract:We have developed our original tissue engineering technology “Cell sheet engineering” utilizing temperature-responsive culture dishes. The Cells are confluently grown on a temperature-responsive culture dish and can be harvested as a Cell sheet by lowering temperature without enzymatic digestion. Cell sheets are high-Cell-density tissues similar to actual living tissues, maintaining their structure and function. Based on this “Cell sheet engineering”, we are trying to create functional Cardiac tissues from human induced pluripotent stem Cells, for regenerative therapy and in vitro drug testing. Toward this purpose, it is necessary to evaluate the contractility of engineered Cardiac Cell sheets. Therefore, in the present study, we developed a contractile force measurement system and evaluated the contractility of human iPSC-derived Cardiac Cell sheet-tissues. By attaching the Cardiac Cell sheets on fibrin gel sheets, we created dynamically beating Cardiac Cell sheet-tissues. They were mounted to the force measurement system and the contractile force was measured stably and clearly. The absolute values of contractile force were around 1 mN, and the mean force value per cross-sectional area was 3.3 mN/mm2. These values are equivalent to or larger than many previously reported values, indicating the functionality of our engineered Cardiac Cell sheets. We also confirmed that both the contractile force and beating rate were significantly increased by the administration of adrenaline, which are the physiologically relevant responses for Cardiac tissues. In conclusion, the force measurement system developed in the present study is valuable for the evaluation of engineered Cardiac Cell sheet-tissues, and for in vitro drug testing as well.
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creation of human Cardiac Cell sheets using pluripotent stem Cells
Biochemical and Biophysical Research Communications, 2012Co-Authors: Katsuhisa Matsuura, Masanori Wada, Tatsuya Shimizu, Yuji Haraguchi, Fumiko Sato, Kasumi Sugiyama, Kanako Konishi, Yuji Shiba, Hinako Ichikawa, Aki TachibanaAbstract:Abstract Although we previously reported the development of Cell-dense thickened Cardiac tissue by repeated transplantation-based vascularization of neonatal rat Cardiac Cell sheets, the Cell sources for human Cardiac Cells sheets and their functions have not been fully elucidated. In this study, we developed a bioreactor to expand and induce Cardiac differentiation of human induced pluripotent stem Cells (hiPSCs). Bioreactor culture for 14 days produced around 8 × 10 7 Cells/100 ml vessel and about 80% of Cells were positive for Cardiac troponin T. After Cardiac differentiation, cardiomyocytes were cultured on temperature-responsive culture dishes and showed spontaneous and synchronous beating, even after Cell sheets were detached from culture dishes. Furthermore, extraCellular action potential propagation was observed between Cell sheets when two Cardiac Cell sheets were partially overlaid. These findings suggest that Cardiac Cell sheets formed by hiPSC-derived cardiomyocytes might have sufficient properties for the creation of thickened Cardiac tissue.
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creation of mouse embryonic stem Cell derived Cardiac Cell sheets
Biomaterials, 2011Co-Authors: Katsuhisa Matsuura, Tatsuya Shimizu, Yuji Haraguchi, Shinako Masuda, Noriko Yasuda, Nobuhisa Hagiwara, Peter W Zandstra, Teruo OkanoAbstract:Abstract Research on heart tissue engineering is an exciting and promising area. Although we previously developed bioengineered myocardium using Cell sheet-based tissue engineering technologies, the issue of appropriate Cell sources remained unresolved. In the present study, we created Cell sheets of mouse embryonic stem (ES) Cell-derived cardiomyocytes after expansion in three-dimensional stirred suspension cultures. Serial treatment of the suspension cultures with noggin and granulocyte colony-stimulating factor significantly increased the number of cardiomyocytes by more than fourfold compared with untreated cultures. After drug selection for ES Cells expressing the neomycin-resistance gene under the control of the α-myosin heavy chain promoter, almost all of the Cells showed spontaneous beating and expressed several Cardiac contractive proteins in a fine striated pattern. When ES-derived cardiomyocytes alone were seeded onto temperature-responsive culture dishes, Cell sheets were not created, whereas cocultures with Cardiac fibroblasts promoted Cell sheet formation. The cardiomyocytes in the Cell sheets beat spontaneously and synchronously, and expressed connexin 43 at the edge of adjacent cardiomyocytes. Furthermore, when the extraCellular action potential was recorded, unidirectional action potential propagation was observed. The present findings suggest that stirred suspension cultures with appropriate growth factors are capable of producing cardiomyocytes effectively and easily, and that ES-derived Cardiac Cell sheets may be a promising tool for the development of bioengineered myocardium.
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Cardiac Cell sheet transplantation improves damaged heart function via superior Cell survival in comparison with dissociated Cell injection
Tissue Engineering Part A, 2011Co-Authors: Hidekazu Sekine, Katsuhisa Matsuura, Tatsuya Shimizu, Masayuki Yamato, Nobuhisa Hagiwara, Izumi Dobashi, Masafumi Takahashi, Eiji Kobayashi, Teruo OkanoAbstract:Regenerative therapies have currently emerged as one of the most promising treatments for repair of the damaged heart. Recently, numerous researchers reported that isolated Cell injection treatments can improve heart function in myocardial infarction models. However, significant Cell loss due to primary hypoxia or Cell wash-out and difficulty to control the location of the grafted Cells remains problem. As an attempt to overcome these limitations, we have proposed Cell sheet-based tissue engineering, which involves stacking confluently cultured Cells (two-dimensional), Cell sheets, to construct three-dimensional Cell-dense tissues. Cell sheet transplantation has been able to recover damaged heart function. However, no detailed analysis for transplanted Cell survival has been previously performed. The present study compared the survival of Cardiac Cell sheet transplantation to direct Cell injection in a rat myocardial infarction model. Luciferase-expressing neonatal rat Cardiac Cells were harvested as Cell...
Smadar Cohen - One of the best experts on this subject based on the ideXlab platform.
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activation of the erk1 2 cascade via pulsatile interstitial fluid flow promotes Cardiac tissue assembly
Tissue Engineering, 2007Co-Authors: Tal Dvir, Oren Levy, Michal Shachar, Yosef Granot, Smadar CohenAbstract:Deciphering the Cellular signals leading to Cardiac muscle assembly is a major challenge in ex vivo tissue regeneration. For the first time, we demonstrate that pulsatile interstitial fluid flow in three-dimensional neonatal Cardiac Cell constructs can activate ERK1/2 sixfold, as compared to static-cultivated constructs. Activation of ERK1/2 was attained under physiological shear stress conditions, without activating the p38 Cell death signal above its basic level. Activation of the ERK1/2 signaling cascade induced synthesis of high levels of contractile and Cell-Cell contact proteins by the cardiomyocytes, while its inhibition diminished the inducing effects of pulsatile flow. The pulsed medium-induced Cardiac Cell constructs showed improved Cellularity and viability, while the regenerated Cardiac tissue demonstrated some ultra-structural features of the adult myocardium. The cardiomyocytes were elongated and aligned into myofibers with defined Z-lines and multiple high-ordered sarcomeres. Numerous inter...
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optimization of Cardiac Cell seeding and distribution in 3d porous alginate scaffolds
Biotechnology and Bioengineering, 2002Co-Authors: Ayelet Dar, Michal Shachar, Jonathan Leor, Smadar CohenAbstract:Cardiac tissue engineering has evolved as a potential therapeutic approach to assist in Cardiac regeneration. We have recently shown that tissue-engineered Cardiac graft, constructed from cardiomyocytes seeded within an alginate scaffold, is capable of preventing the deterioration in Cardiac function after myocardial infarction in rats. The present article addresses Cell seeding within porous alginate scaffolds in an attempt to achieve 3D high-density Cardiac constructs with a uniform Cell distribution. Due to the hydrophilic nature of the alginate scaffold, its >90% porosity and interconnected pore structure, Cell seeding onto the scaffold was efficient and short, up to 30 min. Application of a moderate centrifugal force during Cell seeding resulted in a uniform Cell distribution throughout the alginate scaffolds, consequently enabling the loading of a large number of Cells onto the 3D scaffolds. The percent Cell yield in the alginate scaffolds ranged between 60-90%, depending on Cell density at seeding; it was 90% at seeding densities of up to 1 x 10 8 Cells/cm 3 scaffold and decreased to 60% at higher densities. The highly dense Cardiac constructs maintained high metabolic activity in culture. Scanning electron microscopy revealed that the Cells aggregated within the scaffold pores. Some of the aggregates were contracting spontaneously within the matrix pores. Throughout the culture there was no indication of cardiomyocyte proliferation within the scaffolds, nor was it found in 3D cultures of cardiofibroblasts. This may enable the development of Cardiac cocultures, without domination of cardiofibroblasts with time.
Hidekazu Sekine - One of the best experts on this subject based on the ideXlab platform.
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Engineering of functional Cardiac tubes by stepwise transplantation of Cardiac Cell sheets onto intestinal mesentery
Heart and Vessels, 2020Co-Authors: Noriyasu Masuda, Hidekazu Sekine, Hiroshi Niinami, Tatsuya ShimizuAbstract:Implantable organ-like grafts made using tissue engineering techniques could potentially be used as circulatory assist devices in people with heart failure. The aims of this study were to engineer implantable, thick Cardiac tubes by the stepwise transplantation of Cardiac Cell sheets onto intestinal mesentery and confirm that these Cardiac tubes exhibited pulsatile activity and generated an internal pressure. Cell sheets were created by culturing neonatal rat Cardiac Cells on temperature-responsive dishes. After harvesting, three Cell sheets were stacked, and the triple-layered Cell sheet was rolled around a section of endotracheal tube. The resulting construct was cultured to generate a Cardiac tube. In the single-step group ( n = 6), a Cardiac tube was implanted onto the intestinal mesentery of a rat. In the double-step group ( n = 6), a Cardiac tube was implanted onto the intestinal mesentery of a rat, and another new Cardiac tube was inserted into the original Cardiac tube one day later. The pulsations and internal pressures of the implanted Cardiac tubes were evaluated 1, 2 and 4 weeks after transplantation. Histology and immunohistochemistry were used to confirm whether vasculature was present in the Cardiac tubes at 4 weeks after transplantation. We found that the Cardiac tubes developed spontaneous pulsations from 1 week after transplantation. The average internal pressures of the Cardiac tubes at 4 weeks after transplantation were 1.8 ± 1.0 mmHg in the single-step group and 2.5 ± 0.3 mmHg in the double-step group. The Cardiac tubes in the double-step group contracted in response to electrical stimulation at 4 weeks after transplantation. Histological and immunohistochemical analyses revealed engraftment of the transplanted Cardiac Cell sheets and neovascularization of the Cardiac tubes in both groups. Our findings demonstrate that it is feasible to generate functional Cardiac tubes using the mesentery as a vascular bed. Further development of this technique will include the creation of a thicker tube, transplantation of the tube into major vessels and evaluation of the function of the tube under physiological conditions.
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Cardiac Cell sheet transplantation improves damaged heart function via superior Cell survival in comparison with dissociated Cell injection
Tissue Engineering Part A, 2011Co-Authors: Hidekazu Sekine, Katsuhisa Matsuura, Tatsuya Shimizu, Masayuki Yamato, Nobuhisa Hagiwara, Izumi Dobashi, Masafumi Takahashi, Eiji Kobayashi, Teruo OkanoAbstract:Regenerative therapies have currently emerged as one of the most promising treatments for repair of the damaged heart. Recently, numerous researchers reported that isolated Cell injection treatments can improve heart function in myocardial infarction models. However, significant Cell loss due to primary hypoxia or Cell wash-out and difficulty to control the location of the grafted Cells remains problem. As an attempt to overcome these limitations, we have proposed Cell sheet-based tissue engineering, which involves stacking confluently cultured Cells (two-dimensional), Cell sheets, to construct three-dimensional Cell-dense tissues. Cell sheet transplantation has been able to recover damaged heart function. However, no detailed analysis for transplanted Cell survival has been previously performed. The present study compared the survival of Cardiac Cell sheet transplantation to direct Cell injection in a rat myocardial infarction model. Luciferase-expressing neonatal rat Cardiac Cells were harvested as Cell...
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endothelial Cell coculture within tissue engineered cardiomyocyte sheets enhances neovascularization and improves Cardiac function of ischemic hearts
Circulation, 2008Co-Authors: Hidekazu Sekine, Tatsuya Shimizu, Sachiko Sekiya, Masayuki Yamato, Eiji Kobayashi, Kyoko Hobo, Joseph Yang, Hiromi Kurosawa, Teruo OkanoAbstract:Background— Regenerative therapies, including myocardial tissue engineering, have been pursued as a new possibility to repair the damaged myocardium, and previously the transplantation of layered cardiomyocyte sheets has been shown to be able to improve Cardiac function after myocardial infarction. We examined the effects of promoting neovascularization by controlling the densities of cocultured endothelial Cells (ECs) within engineered myocardial tissues created using our Cell sheet-based tissue engineering approach. Methods and Results— Neonatal rat cardiomyocytes were cocultured with GFP-positive rat-derived ECs on temperature-responsive culture dishes. Cocultured ECs formed Cell networks within the cardiomyocyte sheets, which were preserved during Cell harvest from the dishes using simple temperature reduction. We also observed significantly increased in vitro production of vessel-forming cytokines by the EC-positive Cardiac Cell sheets. After layering of 3 Cardiac Cell sheets to create 3-dimensional ...
Katsuhisa Matsuura - One of the best experts on this subject based on the ideXlab platform.
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Contractile force measurement of human induced pluripotent stem Cell-derived Cardiac Cell sheet-tissue.
PLOS ONE, 2018Co-Authors: Daisuke Sasaki, Katsuhisa Matsuura, Yuji Haraguchi, Hiroyoshi Seta, Teruo Okano, Tatsuya ShimizuAbstract:We have developed our original tissue engineering technology “Cell sheet engineering” utilizing temperature-responsive culture dishes. The Cells are confluently grown on a temperature-responsive culture dish and can be harvested as a Cell sheet by lowering temperature without enzymatic digestion. Cell sheets are high-Cell-density tissues similar to actual living tissues, maintaining their structure and function. Based on this “Cell sheet engineering”, we are trying to create functional Cardiac tissues from human induced pluripotent stem Cells, for regenerative therapy and in vitro drug testing. Toward this purpose, it is necessary to evaluate the contractility of engineered Cardiac Cell sheets. Therefore, in the present study, we developed a contractile force measurement system and evaluated the contractility of human iPSC-derived Cardiac Cell sheet-tissues. By attaching the Cardiac Cell sheets on fibrin gel sheets, we created dynamically beating Cardiac Cell sheet-tissues. They were mounted to the force measurement system and the contractile force was measured stably and clearly. The absolute values of contractile force were around 1 mN, and the mean force value per cross-sectional area was 3.3 mN/mm2. These values are equivalent to or larger than many previously reported values, indicating the functionality of our engineered Cardiac Cell sheets. We also confirmed that both the contractile force and beating rate were significantly increased by the administration of adrenaline, which are the physiologically relevant responses for Cardiac tissues. In conclusion, the force measurement system developed in the present study is valuable for the evaluation of engineered Cardiac Cell sheet-tissues, and for in vitro drug testing as well.
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creation of human Cardiac Cell sheets using pluripotent stem Cells
Biochemical and Biophysical Research Communications, 2012Co-Authors: Katsuhisa Matsuura, Masanori Wada, Tatsuya Shimizu, Yuji Haraguchi, Fumiko Sato, Kasumi Sugiyama, Kanako Konishi, Yuji Shiba, Hinako Ichikawa, Aki TachibanaAbstract:Abstract Although we previously reported the development of Cell-dense thickened Cardiac tissue by repeated transplantation-based vascularization of neonatal rat Cardiac Cell sheets, the Cell sources for human Cardiac Cells sheets and their functions have not been fully elucidated. In this study, we developed a bioreactor to expand and induce Cardiac differentiation of human induced pluripotent stem Cells (hiPSCs). Bioreactor culture for 14 days produced around 8 × 10 7 Cells/100 ml vessel and about 80% of Cells were positive for Cardiac troponin T. After Cardiac differentiation, cardiomyocytes were cultured on temperature-responsive culture dishes and showed spontaneous and synchronous beating, even after Cell sheets were detached from culture dishes. Furthermore, extraCellular action potential propagation was observed between Cell sheets when two Cardiac Cell sheets were partially overlaid. These findings suggest that Cardiac Cell sheets formed by hiPSC-derived cardiomyocytes might have sufficient properties for the creation of thickened Cardiac tissue.
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creation of mouse embryonic stem Cell derived Cardiac Cell sheets
Biomaterials, 2011Co-Authors: Katsuhisa Matsuura, Tatsuya Shimizu, Yuji Haraguchi, Shinako Masuda, Noriko Yasuda, Nobuhisa Hagiwara, Peter W Zandstra, Teruo OkanoAbstract:Abstract Research on heart tissue engineering is an exciting and promising area. Although we previously developed bioengineered myocardium using Cell sheet-based tissue engineering technologies, the issue of appropriate Cell sources remained unresolved. In the present study, we created Cell sheets of mouse embryonic stem (ES) Cell-derived cardiomyocytes after expansion in three-dimensional stirred suspension cultures. Serial treatment of the suspension cultures with noggin and granulocyte colony-stimulating factor significantly increased the number of cardiomyocytes by more than fourfold compared with untreated cultures. After drug selection for ES Cells expressing the neomycin-resistance gene under the control of the α-myosin heavy chain promoter, almost all of the Cells showed spontaneous beating and expressed several Cardiac contractive proteins in a fine striated pattern. When ES-derived cardiomyocytes alone were seeded onto temperature-responsive culture dishes, Cell sheets were not created, whereas cocultures with Cardiac fibroblasts promoted Cell sheet formation. The cardiomyocytes in the Cell sheets beat spontaneously and synchronously, and expressed connexin 43 at the edge of adjacent cardiomyocytes. Furthermore, when the extraCellular action potential was recorded, unidirectional action potential propagation was observed. The present findings suggest that stirred suspension cultures with appropriate growth factors are capable of producing cardiomyocytes effectively and easily, and that ES-derived Cardiac Cell sheets may be a promising tool for the development of bioengineered myocardium.
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Cardiac Cell sheet transplantation improves damaged heart function via superior Cell survival in comparison with dissociated Cell injection
Tissue Engineering Part A, 2011Co-Authors: Hidekazu Sekine, Katsuhisa Matsuura, Tatsuya Shimizu, Masayuki Yamato, Nobuhisa Hagiwara, Izumi Dobashi, Masafumi Takahashi, Eiji Kobayashi, Teruo OkanoAbstract:Regenerative therapies have currently emerged as one of the most promising treatments for repair of the damaged heart. Recently, numerous researchers reported that isolated Cell injection treatments can improve heart function in myocardial infarction models. However, significant Cell loss due to primary hypoxia or Cell wash-out and difficulty to control the location of the grafted Cells remains problem. As an attempt to overcome these limitations, we have proposed Cell sheet-based tissue engineering, which involves stacking confluently cultured Cells (two-dimensional), Cell sheets, to construct three-dimensional Cell-dense tissues. Cell sheet transplantation has been able to recover damaged heart function. However, no detailed analysis for transplanted Cell survival has been previously performed. The present study compared the survival of Cardiac Cell sheet transplantation to direct Cell injection in a rat myocardial infarction model. Luciferase-expressing neonatal rat Cardiac Cells were harvested as Cell...