2017 Fiscal Year Research-status Report
High efficient myogenic differentiation of human pluripotent stem cells on New Generation Laminins towards regenerative medicine.
Project/Area Number |
17K13014
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Research Institution | Kyoto University |
Principal Investigator |
Zhao Mingming 京都大学, iPS細胞研究所, 特定研究員 (50754206)
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Project Period (FY) |
2017-04-01 – 2019-03-31
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Keywords | 再生医学 / 次世代ラミニン / iPS細胞 / 骨格筋幹細胞 |
Outline of Annual Research Achievements |
Using NGL (New Generation Laminin), we established a highly efficient differentiation system for myocytes and muscle stem cells induction from hiPSCs for disease modeling and cell therapy. Instead of animal derived Matrigel, the NGL promotes the development of paraxial mesoderm, dermomyotome, myogenic progenitors and muscle stem cells from hiPSCs in the stepwise differentiation protocol. The results demonstrated that the unique crystal structure of NGL supports the high efficient myogenic differentiation through FGF receptor signaling pathway regulated by heparan sulfates. Recent studies are focusing on the mechanisms of NGL regulating myogenic differentiation in hiPSCs.
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Current Status of Research Progress |
Current Status of Research Progress
1: Research has progressed more than it was originally planned.
Reason
By screening all of the NGL (New Generation Laminin) isoforms which could be obtained, we revealed that p421 provided the most efficient myogenic differentiation. Using p421 we successfully established a more efficient differentiation system for myocytes and muscle stem cells induction from hiPSCs. The Heparan sulfates digestion experiment revealed that the unique effects of p421 is provided by HS (Heparan sulfate). By screening the pathways of FGF, WNT, BMP, TGFβ, PDGF signaling, we revealed that p421 regulating myogenic differentiation by regulating FGFR signaling pathway. Further evaluating the relationship of p421 and FGFR signaling pathway is currently in progress.
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Strategy for Future Research Activity |
Depend on the results we already received, we will continue to elucidate the mechanism of HS in p421 regulating FGFR signaling pathway. shRNA Knock down system will be performed to confirm the function of FGF, FGFR or specific integrin in our system. Following our initial strategy, we will continue to establish the strategy to purify iPSCs derived myocytes and muscle stem cells, then confirm the characterization of myocytes and muscle stem cells in vitro and in vivo.
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Causes of Carryover |
Incurring amount will be used for the reagent for cell culture, shRNA kit, antibody and other experiment consumables which will be acquired, and for domestic and overseas travel expenses. The reagents and consumables which will be acquired for this project will be used in accordance with our stipulated research plan in a timely and efficient manner.
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Research Products
(4 results)