title: [Generation of a P19-alphaMHC-EGFP reporter line and cardiomyocyte differentiation].
abstract: OBJECTIVE
  To generate a P19-alphaMHC-EGFP reporter line and induce cardiomyocyte differentiation of this reporter line.
  
  
  METHODS
  The P19 cells were transfected with palphaMHC-EGFP, a P19-alphaMHC-EGFP reporter line was obtained after G418 selection and limited dilution of recombinant clones. The reporter line was induced to differentiate into cardiomyocytes which would beat and express green fluorescent protein. A comparison of cardiomyocyte differentiation rate and cTnI expression amount between the reporter line and the untransfected P19 cells was also performed. The ultrastructure was observed under transmission electron microscope.
  
  
  RESULTS
  The ultrastructure characteristics indicated cardiomyocytes-like changes on induction day 10. The beating cardiomyocytes which express GFP appear in the seventh induction day. The cardiomyocyte differentiation rate and cTnI expression amount of P19-alphaMHC-EGFP reporter line were similar as those in untransfected P19 cells (P > 0.05).
  
  
  CONCLUSIONS
  The P19-alphaMHC-EGFP reporter line is of great benefit for identifying and purifying cardiomyocytes from undifferentiated P19 cells without influencing the differentiation of P19 cells. This feature makes P19-alphaMHC-EGFP reporter line a promising cell source for clinical cardiomyocyte replacement therapy.
authors.affiliation: Department of Cardiology, First Affiliated Hospital, Nanjing Medical University, Cardiovascular Medical Center of Jiangsu, Nanjing 210029, China.
authors.forename: Jie
authors.initials: J
authors.lastname: Gong
authors.forename: Qi-jun
authors.initials: QJ
authors.lastname: Zhang
authors.forename: Jian
authors.initials: J
authors.lastname: Wang
authors.forename: Feng-rong
authors.initials: FR
authors.lastname: Sun
authors.forename: Ling-mei
authors.initials: LM
authors.lastname: Qian
authors.forename: Xiang-qing
authors.initials: XQ
authors.lastname: Kong
authors.forename: Rong
authors.initials: R
authors.lastname: Yang
authors.forename: Yan-hui
authors.initials: YH
authors.lastname: Sheng
authors.forename: Ke-jiang
authors.initials: KJ
authors.lastname: Cao
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002460:Cell Line; D051379:Mice; D032383:Myocytes, Cardiac / Q000166:cytology*; D013234:Stem Cells / Q000166:cytology*; D014162:Transfection