title: Differentiation of adipose tissue-derived mesenchymal stem cells into cardiomyocytes.
abstract: BACKGROUND
  Cardiomyocytes have small potential for renovation and proliferation in vivo. Consequently, the heart muscle has limited capacity of self-renewal. Mesenchymal stem cells (MSC) therapy, as well as MSC differentiated into cardiomyocytes, has been used in the attempt to minimize the effects of ischemic-hypoxic lesions and those affecting the electrical conduction system of the heart.
  
  
  OBJECTIVE
  The present study compared three distinct protocols for induced differentiation of MSC into cardiomyocytes aimed at finding a viable method for producing a large number of functional cells expressing cardiomyogenic phenotype.
  
  
  METHODS
  Mesenchymal stem cells were obtained from the adipose tissue of young transgenic Lewis rats expressing green fluorescent protein (GFP), and submitted to three distinct differentiation-inducing media: 1) Planat-Bérnard, 2) 5-azacytidine, and 3) Planat-Bérnard + 5-azacytidine; further, these cells were identified based on the expression of cardiac cell markers.
  
  
  RESULTS
  All three protocols detected the expression of sarcomeric-alpha-actinin protein in the exoskeleton of cells, expression of connexin-43 in the nuclear and cytoplasmic membrane, and formation of gap junctions, which are necessary for electrical impulse propagation in the myocardium. However, no spontaneous cell contraction was observed with any of the tested protocols.
  
  
  CONCLUSIONS
  Induction with 5-azacytidine provided an effective cadiomyogenic cellular differentiation similar to that obtained with Planat-Bénard media. Therefore, 5-azacytidine was the method of choice for being the simplest, fastest and lowest-cost protocol for cell differentiation.
authors.affiliation: Universidade Federal de Viçosa, Viçosa, MG, Brasil.
authors.forename: Pablo Herthel
authors.initials: PH
authors.lastname: Carvalho
authors.forename: Ana Paula Falci
authors.initials: AP
authors.lastname: Daibert
authors.forename: Betânia Souza
authors.initials: BS
authors.lastname: Monteiro
authors.forename: Bárbara Silva
authors.initials: BS
authors.lastname: Okano
authors.forename: Juliana Lott
authors.initials: JL
authors.lastname: Carvalho
authors.forename: Daise Nunes Queiroz da
authors.initials: DN
authors.lastname: Cunha
authors.forename: Lukiya Silva Campos
authors.initials: LS
authors.lastname: Favarato
authors.forename: Vanessa Guedes
authors.initials: VG
authors.lastname: Pereira
authors.forename: Luis Eugênio Franklin
authors.initials: LE
authors.lastname: Augusto
authors.forename: Ricardo Junqueira
authors.initials: RJ
authors.lastname: Del Carlo
mesh_terms: D017667:Adipocytes / Q000166:cytology* / Q000187:drug effects; D000273:Adipose Tissue / Q000166:cytology*; D000818:Animals; D001374:Azacitidine / Q000494:pharmacology; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D032383:Myocytes, Cardiac / Q000166:cytology*; D051381:Rats; D011917:Rats, Inbred Lew; D015203:Reproducibility of Results
chemical_list: D001374:Azacitidine