| Autor: |
Maffioletti SM; Department of Cell and Developmental Biology, University College London, London, UK., Gerli MF; Department of Cell and Developmental Biology, University College London, London, UK., Ragazzi M; Department of Cell and Developmental Biology, University College London, London, UK., Dastidar S; Department of Gene Therapy and Regenerative Medicine, Free University of Brussels (VUB), Brussels, Belgium., Benedetti S; 1] Department of Cell and Developmental Biology, University College London, London, UK. [2] Present address: Institute of Child Health, University College London, London, UK., Loperfido M; 1] Department of Gene Therapy and Regenerative Medicine, Free University of Brussels (VUB), Brussels, Belgium. [2] Department of Cardiovascular Sciences, Center for Molecular and Vascular Biology, University of Leuven (KU Leuven), Leuven, Belgium., VandenDriessche T; 1] Department of Gene Therapy and Regenerative Medicine, Free University of Brussels (VUB), Brussels, Belgium. [2] Department of Cardiovascular Sciences, Center for Molecular and Vascular Biology, University of Leuven (KU Leuven), Leuven, Belgium., Chuah MK; 1] Department of Gene Therapy and Regenerative Medicine, Free University of Brussels (VUB), Brussels, Belgium. [2] Department of Cardiovascular Sciences, Center for Molecular and Vascular Biology, University of Leuven (KU Leuven), Leuven, Belgium., Tedesco FS; Department of Cell and Developmental Biology, University College London, London, UK. |
| Abstrakt: |
Skeletal muscle is the most abundant human tissue; therefore, an unlimited availability of myogenic cells has applications in regenerative medicine and drug development. Here we detail a protocol to derive myogenic cells from human embryonic stem (ES) and induced pluripotent stem (iPS) cells, and we also provide evidence for its extension to human iPS cells cultured without feeder cells. The procedure, which does not require the generation of embryoid bodies or prospective cell isolation, entails four stages with different culture densities, media and surface coating. Pluripotent stem cells are disaggregated to single cells and then differentiated into expandable cells resembling human mesoangioblasts. Subsequently, transient Myod1 induction efficiently drives myogenic differentiation into multinucleated myotubes. Cells derived from patients with muscular dystrophy and differentiated using this protocol have been genetically corrected, and they were proven to have therapeutic potential in dystrophic mice. Thus, this platform has been demonstrated to be amenable to gene and cell therapy, and it could be extended to muscle tissue engineering and disease modeling. |
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