Derivation of Skeletal Myogenic Precursors from Human Pluripotent Stem Cells Using Conditional Expression of PAX7.
Cell-based therapies are considered as one of the most promising approaches for the treatment of degenerating pathologies including muscle disorders and dystrophies. Advances in the approach of reprogramming somatic cells into induced pluripotent stem (iPS) cells allow for the possibility of using the patient's own pluripotent cells to generate specific tissues for autologous transplantation. In addition, patient-specific tissue derivatives have been shown to represent valuable material for disease modeling and drug discovery. Nevertheless, directed differentiation of pluripotent stem cells into a specific lineage is not a trivial task especially in the case of skeletal myogenesis, which is generally poorly recapitulated during the in vitro differentiation of pluripotent stem cells.Here, we describe a practical and efficient method for the derivation of skeletal myogenic precursors from differentiating human pluripotent stem cells using controlled expression of PAX7. Flow cytometry (FACS) purified myogenic precursors can be expanded exponentially and differentiated in vitro into myotubes, enabling researchers to use these cells for disease modeling as well as therapeutic purposes.
Center for Stem Cell and Regenerative Medicine/The Brown Foundation Institute of Molecular Medicine, The University of Texas Health Science Center at Houston, 1825 Pressler Street SRB 630A, Houston, TX, 77030, USA.
Radbod
R
Darabi
Department of Medicine, Lillehei Heart Institute, University of Minnesota, CCRB 4-128, 2231 6th St. SE, Minneapolis, MN, 55455, USA. Perli032@umn.edu.
Rita C R
RC
Perlingeiro
D018929:Cell Culture Techniques / Q000379:methods*; D002455:Cell Division; D002469:Cell Separation; D064987:Cell- and Tissue-Based Therapy; D002478:Cells, Cultured; D065150:Cellular Reprogramming; D000067470:Cellular Reprogramming Techniques / Q000379:methods*; D004318:Doxycycline / Q000494:pharmacology; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D005434:Flow Cytometry; D015870:Gene Expression / Q000187:drug effects; D017930:Genes, Reporter; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics; D057809:HEK293 Cells; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism; D016086:Lentivirus / Q000235:genetics; D024510:Muscle Development; D018485:Muscle Fibers, Skeletal / Q000166:cytology*; D051763:PAX7 Transcription Factor / Q000096:biosynthesis* / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D020218:Response Elements / Q000235:genetics; D014161:Transduction, Genetic; D019076:Transgenes
D051763:PAX7 Transcription Factor; C497321:PAX7 protein, human; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins; D004318:Doxycycline
Cell therapy; Human embryonic stem (ES) cells; Human-induced pluripotent stem (iPS) cells; Muscle differentiation; Myogenic precursors; PAX7; Skeletal muscle