Enhanced Cardiomyogenic Differentiation of P19 Embryonal Carcinoma Stem Cells.
10.4070/kcj.2009.39.5.198
- Author:
Jihyun YANG
1
;
Seok Jin KO
;
Beom Suk KIM
;
Hyun Seo KIM
;
Sangheon PARK
;
Doran HONG
;
Soon Woong HONG
;
Ji Hyun CHOI
;
Chi Yeon PARK
;
Seung Cheol CHOI
;
Sun Jun HONG
;
Do Sun LIM
Author Information
1. Department of Cardiology, College of Medicine, Korea University, Seoul, Korea. dslmd@kumc.or.kr
- Publication Type:Original Article
- Keywords:
Cell differentiation;
Embryonal carcinoma stem cells;
Myocytes, cardiac;
Serum
- MeSH:
Actins;
Antibodies;
Azacitidine;
Carcinoma, Embryonal;
Cell Differentiation;
Dimethyl Sulfoxide;
Embryoid Bodies;
Embryonal Carcinoma Stem Cells;
Myocytes, Cardiac;
Real-Time Polymerase Chain Reaction;
RNA;
Safrole;
Troponin T;
Ventricular Myosins
- From:Korean Circulation Journal
2009;39(5):198-204
- CountryRepublic of Korea
- Language:English
-
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 AND METHODS: 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.
