[Generation of a P19-alphaMHC-EGFP reporter line and cardiomyocyte differentiation].
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.
Gong
Jie
J
Department of Cardiology, First Affiliated Hospital, Nanjing Medical University, Cardiovascular Medical Center of Jiangsu, Nanjing 210029, China.
Zhang
Qi-jun
QJ
Wang
Jian
J
Sun
Feng-rong
FR
Qian
Ling-mei
LM
Kong
Xiang-qing
XQ
Yang
Rong
R
Sheng
Yan-hui
YH
Cao
Ke-jiang
KJ
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