Use of Long-term Cultured Embryoid Bodies May Enhance Cardiomyocyte Differentiation by BMP2.
10.3349/ymj.2008.49.5.819
- Author:
Yoon Young KIM
1
;
Seung Yup KU
;
Jiho JANG
;
Sun Kyung OH
;
Hee Sun KIM
;
Seok Hyun KIM
;
Young Min CHOI
;
Shin Yong MOON
Author Information
1. Institute of Reproductive Medicine and Population, Medical Research Center, Seoul, Korea. ymchoi@snu.ac.kr
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- Keywords:
Bone morphogenetic protein 2;
cardiomyocytes;
cell differentiation;
embryoid bodies;
embryonic stem cells;
long-term
- MeSH:
Bone Morphogenetic Protein 2/*pharmacology;
Cell Culture Techniques;
*Cell Differentiation;
Cell Line;
Cell Proliferation;
Embryonic Stem Cells/cytology/*drug effects;
Humans;
Myocytes, Cardiac/*cytology;
Pluripotent Stem Cells/cytology/drug effects;
Signal Transduction
- From:Yonsei Medical Journal
2008;49(5):819-827
- CountryRepublic of Korea
- Language:English
-
Abstract:
PURPOSE: Human embryonic stem cells (hESCs) can proliferate for a prolonged period and differentiate into cardiomyocytes in vitro. Recent studies used bone morphogenetic protein 2 (BMP2) to generate cardiomyocytes from hESCs, however, all those studies used early embryoid bodies (EBs) and did not retrieve cardiomyocytes with a high yield. In this study, we treated long-term cultured EBs with BMP2 in order to promote differentiation into cardiomyocytes from hESCs. MATERIALS AND METHODS: hESC lines, including SNUhES3 and SNUhES4, were used in this study. Undifferentiated hESC colonies were detached to form EBs and cultured for up to 30 days. These long-term cultured EBs were differentiated into cardiomyocytes in serum-containing media. In our protocol, BMP2 was applied for 5 days after attachment of EBs. Cardiac specific markers, beating of differentiated cells and electron microscopic (EM) ultrastructures were evaluated and analyzed. RESULTS: Compared to 10-day or 20-day EBs, 30-day EBs showed a higher expression level of cardiac specific markers, Nkx2.5 and a-myosin heavy chain (alphaMHC). Treatment of BMP2 increased expression of cardiac troponin (cTn) I and a-actinin when evaluated at 20 days after attachment of 30-day EBs. Beating of differentiated cells was observed from 7 to 20 days after attachment. Moreover, EM findings demonstrated fine structures such as Z bands in these differentiated cardiomyocytes. These long-term cultured EBs yielded cardiomyocytes with an efficiency of as high as 73.6% when assessed by FACS. CONCLUSION: We demonstrated that the use of long-term cultured EBs may enhance differentiation into cardiomyocytes from hESCs when treated with BMP2.
