Isolation and enrichment of skeletal muscle progenitor cells from mouse bone marrow.
There is great interest in the therapeutic potential of non-hematopoietic stem cells obtained from bone marrow called mesenchymal stem cells (MSCs). Rare myogenic progenitor cells in MSC cultures have been shown to convert into skeletal muscle cells in vitro and also in vivo after transplantation of bone marrow into mice. To be clinically useful, however, isolation and expansion of myogenic progenitor cells is important to improve the efficacy of cell transplantation in generating normal skeletal muscle cells. We introduced into MSCs obtained from mouse bone marrow, a plasmid vector in which an antibiotic (Zeocin) resistance gene is driven by MyoD and Myf5 enhancer elements, which are selectively active in skeletal muscle progenitor cells. Myogenic precursor cells were then isolated by antibiotic selection, expanded in culture, and shown to differentiate appropriately into multinucleate myotubes in vitro. Our results show that using a genetic selection strategy, an enriched population of myogenic progenitor cells, which will be useful for cell transplantation therapies, can be isolated from MSCs.
Bhagavati
Satyakam
S
Department of Neurology, SUNY Downstate Medical Center, 450 Clarkson Avenue, Brooklyn, NY 11203, USA. satyakamb@hotmail.com
Xu
Weimin
W
D000818:Animals; D001761:Bleomycin / Q000494:pharmacology; D001854:Bone Marrow Cells / Q000166:cytology*; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation; D019070:Cell Lineage; D002469:Cell Separation / Q000379:methods; D002478:Cells, Cultured; D004268:DNA-Binding Proteins; D003893:Desmin / Q000096:biosynthesis / Q000235:genetics; D004351:Drug Resistance / Q000235:genetics; D015870:Gene Expression; D005819:Genetic Markers; D005822:Genetic Vectors; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000648:ultrastructure; D051379:Mice; D008807:Mice, Inbred BALB C; D018485:Muscle Fibers, Skeletal / Q000378:metabolism; D009124:Muscle Proteins / Q000096:biosynthesis / Q000235:genetics; D018482:Muscle, Skeletal / Q000166:cytology*; D032446:Myoblasts / Q000648:ultrastructure; D051787:Myogenic Regulatory Factor 5; D018006:Myogenic Regulatory Factors / Q000096:biosynthesis / Q000235:genetics; D013234:Stem Cells / Q000166:cytology*; D015534:Trans-Activators
D004268:DNA-Binding Proteins; D003893:Desmin; D005819:Genetic Markers; D009124:Muscle Proteins; C497418:Myf5 protein, mouse; D051787:Myogenic Regulatory Factor 5; D018006:Myogenic Regulatory Factors; D015534:Trans-Activators; D001761:Bleomycin; C105427:Zeocin