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 1 × 10 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 μM 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, α-actin, α-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 μM 5-aza-cytidine 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.
| Original language | English |
|---|---|
| Pages (from-to) | 198-204 |
| Number of pages | 7 |
| Journal | Korean Circulation Journal |
| Volume | 39 |
| Issue number | 5 |
| DOIs | |
| Publication status | Published - 2009 May 1 |
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Keywords
- Cell differentiation
- Embryonal carcinoma stem cells
- Myocytes, cardiac
- Serum
ASJC Scopus subject areas
- Cardiology and Cardiovascular Medicine
- Internal Medicine
Cite this
Enhanced cardiomyogenic differentiation of P19 embryonal carcinoma stem cells. / Yang, Jihyun; Ko, Seok Jin; Kim, Beom Suk; Kim, Hyun Seo; Park, Sangheon; Hong, Doran; Hong, Soon Woong; Choi, Ji Hyun; Park, Chi Yeon; Choi, Seung Cheol; Hong, Sun Jun; Lim, Do-Sun.
In: Korean Circulation Journal, Vol. 39, No. 5, 01.05.2009, p. 198-204.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Enhanced cardiomyogenic differentiation of P19 embryonal carcinoma stem cells
AU - Yang, Jihyun
AU - Ko, Seok Jin
AU - Kim, Beom Suk
AU - Kim, Hyun Seo
AU - Park, Sangheon
AU - Hong, Doran
AU - Hong, Soon Woong
AU - Choi, Ji Hyun
AU - Park, Chi Yeon
AU - Choi, Seung Cheol
AU - Hong, Sun Jun
AU - Lim, Do-Sun
PY - 2009/5/1
Y1 - 2009/5/1
N2 - 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 1 × 10 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 μM 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, α-actin, α-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 μM 5-aza-cytidine 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.
AB - 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 1 × 10 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 μM 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, α-actin, α-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 μM 5-aza-cytidine 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.
KW - Cell differentiation
KW - Embryonal carcinoma stem cells
KW - Myocytes, cardiac
KW - Serum
UR - http://www.scopus.com/inward/record.url?scp=67549119402&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=67549119402&partnerID=8YFLogxK
U2 - 10.4070/kcj.2009.39.5.198
DO - 10.4070/kcj.2009.39.5.198
M3 - Article
C2 - 19949579
AN - SCOPUS:67549119402
VL - 39
SP - 198
EP - 204
JO - Korean Circulation Journal
JF - Korean Circulation Journal
SN - 1738-5520
IS - 5
ER -