title: [Human bone marrow mesenchymal stem cells cocultured with semi-permeable membrane separated neonatal rat ventricular myocytes differentiated into cardiomyocyte phenotype].
abstract: OBJECTIVE
  To investigate the ability of human bone marrow mesenchymal stem cells (hBMSCs), cocultured with semi-permeable membrane separated neonatal rat ventricular myocytes, to differentiate into cardiomyocytes.
  
  
  METHODS
  hBMSCs were isolated and purified by density gradient centrifugation and adherence screening method. Cells were expanded as undifferentiated cells in culture for more than 3 passages and their phenotypes were identified with flow cytometer. hBMSCs were cocultured with neonatal rat ventricular myocytes in a rate of 1:10 separated by semi-permeable membrane. GATA4 mRNA was detected by RT-PCR; Immunocytochemistry, and Immunostaining were used to detect sarcomeric alpha-actinin, desmin, cTnT, and cTnI protein level.
  
  
  RESULTS
  CD29 (98.64% +/- 0.80%) and CD44 (96.70% +/- 1.50%) were the major surface markers of hBMSCs. After coculturing with semi-permeable membrane separated neonatal rat ventricular myocytes, the first contraction of single cells was noted at day 7 and GATA4 expression was detected on these cells by RT-PCR after 1 to 3 weeks coculture. Desmin, sarcomeric alpha-actinin, cTnI and cTnT could be detected by immunocytochemistry and immunostaining on some of these cells.
  
  
  CONCLUSIONS
  hBMSCs possess the potential to differentiate into myocardial cell phenotype in the cardiac microenvironment. Direct contact with cardiomyocytes was not necessary required for hBMSCs differentiation.
authors.affiliation: Research Center of Medical Sciences, Guangdong Provincial People's Hospital, Guangzhou 510080, China.
authors.forename: Xiao-hong
authors.initials: XH
authors.lastname: Li
authors.forename: Xi-yong
authors.initials: XY
authors.lastname: Yu
authors.forename: Qiu-xiong
authors.initials: QX
authors.lastname: Lin
authors.forename: Zhi-xin
authors.initials: ZX
authors.lastname: Shan
authors.forename: Guang-feng
authors.initials: GF
authors.lastname: Zhang
authors.forename: Xiao
authors.initials: X
authors.lastname: Zhang
authors.forename: Chun-yu
authors.initials: CY
authors.lastname: Deng
authors.forename: Yong-heng
authors.initials: YH
authors.lastname: Fu
authors.forename: Su-juan
authors.initials: SJ
authors.lastname: Kuang
authors.forename: Wei
authors.initials: W
authors.lastname: Huang
authors.forename: Min
authors.initials: M
authors.lastname: Yang
authors.forename: Shu-guang
authors.initials: SG
authors.lastname: Lin
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002478:Cells, Cultured; D018920:Coculture Techniques; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D032383:Myocytes, Cardiac / Q000166:cytology*; D051381:Rats; D017207:Rats, Sprague-Dawley