Enhanced Cardiomyogenic Differentiation of P19 Embryonal Carcinoma Stem Cells
Korean Circulation Journal
;
: 198-204, 2009.
Article
in English
| WPRIM
| ID: wpr-100654
ABSTRACT
BACKGROUND AND OBJECTIVES:
We investigated the effects of different concentrations of serum, 5-azacytidine, and culture time on the cardiomyogenic differentiation of P19 embryonal carcinoma stem cells in the course of developing an efficient protocol for generating the cardiomyogenic lineage. MATERIALS ANDMETHODS:
P19 cells were plated at a density of 1x10(6) cells on 10-cm bacterial dishes for 96 hours in the presence of 1% dimethyl sulfoxide to form embryoid bodies. The embryoid bodies were cultured in medium with 2% or 10% fetal bovine serum for an additional 10 or 15 consecutive days in the presence of 0, 1, or 3 microM 5-azacytidine.RESULTS:
Quantitative real-time polymerase chain reaction (PCR) analysis showed that the messenger ribonucleic acid (mRNA) expression of cardiac muscle-specific genes, such as GATA4, alpha-actin, alpha-myosin heavy chain, and cardiac troponin T, were significantly higher in the 15-day culture groups than in the 10-day culture groups. Furthermore, the cardiac muscle-specific genes were expressed more in the high-serum groups compared to the low-serum groups regardless of the culture time. Cardiomyogenic differentiation of the P19 cells was most effective in 1 microM 5-azacytidine regardless of the serum concentrations. In addition, the stimulation effects of 5-azacytidine on cardiomyogenic differentiation were more significant under low-serum culture conditions compared to high-serum culture conditions. Cardiomyogenic differentiation of P19 cells was further confirmed by immunostaining with cardiac muscle-specific antibodies.CONCLUSION:
Taken together, these results demonstrated that cardiomyogenic differentiation of P19 cells was enhanced by a combination of different experimental factors.
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Safrole
/
Azacitidine
/
RNA
/
Cell Differentiation
/
Dimethyl Sulfoxide
/
Actins
/
Carcinoma, Embryonal
/
Troponin T
/
Ventricular Myosins
/
Myocytes, Cardiac
Language:
English
Journal:
Korean Circulation Journal
Year:
2009
Type:
Article
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