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Simple derivation of skeletal muscle from human pluripotent stem cells using temperature-sensitive Sendai virus vector |
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| Authors: | Ghee Wan Tan Takayuki Kondo Keiko Imamura Mika Suga Takako Enami Ayako Nagahashi Kayoko Tsukita Ikuyo Inoue Jitsutaro Kawaguchi Tsugumine Shu Haruhisa Inoue |
| Affiliation: | 1. Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan Contribution: Data curation (equal), Formal analysis (equal), Validation (equal), Writing - original draft (equal);2. Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan;3. Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan iPSC-based Drug Discovery and Development Team, RIKEN BioResource Research Center (BRC), Kyoto, Japan Medical-risk Avoidance based on iPS Cells Team, RIKEN Center for Advanced Intelligence Project (AIP), Kyoto, Japan Contribution: Supervision (supporting);4. Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan iPSC-based Drug Discovery and Development Team, RIKEN BioResource Research Center (BRC), Kyoto, Japan Contribution: Supervision (supporting), Writing - original draft (supporting);5. Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan Medical-risk Avoidance based on iPS Cells Team, RIKEN Center for Advanced Intelligence Project (AIP), Kyoto, Japan Contribution: Resources (supporting);6. Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan Medical-risk Avoidance based on iPS Cells Team, RIKEN Center for Advanced Intelligence Project (AIP), Kyoto, Japan Contribution: Visualization (supporting);7. Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan iPSC-based Drug Discovery and Development Team, RIKEN BioResource Research Center (BRC), Kyoto, Japan Contribution: Validation (supporting);8. Center for iPS Cell Research and Application (CiRA), Kyoto University, Kyoto, Japan Medical-risk Avoidance based on iPS Cells Team, RIKEN Center for Advanced Intelligence Project (AIP), Kyoto, Japan Contribution: Validation (supporting);9. R&D Center, ID Pharma Co., Ltd, Tsukuba, Japan Contribution: Resources (equal) |
| Abstract: | Human pluripotent stem cells have the potential to differentiate into various cell types including skeletal muscles (SkM), and they are applied to regenerative medicine or in vitro modelling for intractable diseases. A simple differentiation method is required for SkM cells to accelerate neuromuscular disease studies. Here, we established a simple method to convert human pluripotent stem cells into SkM cells by using temperature-sensitive Sendai virus (SeV) vector encoding myoblast determination protein 1 (SeV-Myod1), a myogenic master transcription factor. SeV-Myod1 treatment converted human embryonic stem cells (ESCs) into SkM cells, which expressed SkM markers including myosin heavy chain (MHC). We then removed the SeV vector by temporal treatment at a high temperature of 38℃, which also accelerated mesodermal differentiation, and found that SkM cells exhibited fibre-like morphology. Finally, after removal of the residual human ESCs by pluripotent stem cell-targeting delivery of cytotoxic compound, we generated SkM cells with 80% MHC positivity and responsiveness to electrical stimulation. This simple method for myogenic differentiation was applicable to human-induced pluripotent stem cells and will be beneficial for investigations of disease mechanisms and drug discovery in the future. |
| Keywords: | differentiation method disease modelling high temperature treatment human embryonic stem cells human-induced pluripotent stem cells Myod1 Sendai virus skeletal muscle |