Use of Long-term Cultured Embryoid Bodies May Enhance Cardiomyocyte Differentiation by BMP2
Yonsei Medical Journal
; : 819-827, 2008.
Article
em En
| WPRIM
| ID: wpr-153694
Biblioteca responsável:
WPRO
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.
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Base de dados:
WPRIM
Assunto principal:
Transdução de Sinais
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Diferenciação Celular
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Linhagem Celular
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Técnicas de Cultura de Células
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Miócitos Cardíacos
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Células-Tronco Pluripotentes
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Proliferação de Células
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Células-Tronco Embrionárias
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Proteína Morfogenética Óssea 2
Tipo de estudo:
Guideline
Limite:
Humans
Idioma:
En
Ano de publicação:
2008
Tipo de documento:
Article