title: Efficient non-viral reprogramming of myoblasts to stemness with a single small molecule to generate cardiac progenitor cells.
abstract: The current protocols for generation of induced pluripotent stem (iPS) cells involve genome integrating viral vectors which may induce tumorgenesis. The aim of this study was to develop and optimize a non-viral method without genetic manipulation for reprogramming of skeletal myoblasts (SMs) using small molecules.
  
  
  RESULTS
  SMs from young male Oct3/4-GFP(+) transgenic mouse were treated with DNA methyltransferase (DNMT) inhibitor, RG108. Two weeks later, GFP(+) colonies of SM derived iPS cells (SiPS) expressing GFP and with morphological similarity of mouse embryonic stem (ESCs) were formed and propagated in vitro. SiPS were positive for alkaline phosphatase activity, expressed SSEA1, displayed ES cell specific pluripotency markers and formed teratoma in nude mice. Optimization of culture conditions for embryoid body (EBs) formation yielded spontaneously contracting EBs having morphological, molecular, and ultra-structural similarities with cardiomyocytes and expressed early and late cardiac markers. miR profiling showed abrogation of let-7 family and upregulation of ESCs specific miR-290-295 cluster thus indicating that SiPS were similar to ESCs in miR profile. Four weeks after transplantation into the immunocompetent mice model of acute myocardial infarction (n = 12 per group), extensive myogenesis was observed in SiPS transplanted hearts as compared to DMEM controls (n = 6 per group). A significant reduction in fibrosis and improvement in global heart function in the hearts transplanted with SiPS derived cardiac progenitor cells were observed.
  
  
  CONCLUSIONS
  Reprogramming of SMs by DNMT inhibitor is a simple, reproducible and efficient technique more likely to generate transgene integration-free iPS cells. Cardiac progenitors derived from iPS cells propagated extensively in the infarcted myocardium without tumorgenesis and improved cardiac function.
authors: Department of Pathology, University of Cincinnati, Cincinnati, Ohio, United States of America.
authors: Zeeshan
authors: Z
authors: Pasha
authors: Husnain Kh
authors: HKh
authors: Haider
authors: Muhammad
authors: M
authors: Ashraf
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics; D065150:Cellular Reprogramming / Q000187:drug effects*; D000076123:DNA (Cytosine-5-)-Methyltransferase 1; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000037:antagonists & inhibitors / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D006334:Heart Function Tests / Q000187:drug effects; D007211:Indoles / Q000494:pharmacology*; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism / Q000648:ultrastructure; D008297:Male; D051379:Mice; D035683:MicroRNAs / Q000235:genetics / Q000378:metabolism; D032448:Myoblasts, Skeletal / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D009200:Myocardial Contraction / Q000187:drug effects; D009203:Myocardial Infarction / Q000503:physiopathology / Q000628:therapy; D009206:Myocardium / Q000166:cytology* / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D010797:Phthalimides; D011422:Propionates / Q000494:pharmacology*; D033581:Stem Cell Transplantation; D013724:Teratoma / Q000473:pathology; D014364:Tryptophan / Q000031:analogs & derivatives; D020257:Ventricular Remodeling / Q000187:drug effects; D014780:Viruses / Q000378:metabolism
chemical_list: D015415:Biomarkers; D007211:Indoles; D035683:MicroRNAs; D010797:Phthalimides; D011422:Propionates; C503639:RG108; D014364:Tryptophan; D000076123:DNA (Cytosine-5-)-Methyltransferase 1; D004248:DNA (Cytosine-5-)-Methyltransferases