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Use of long-term cultured embryoid bodies may enhance cardiomyocyte differentiation by BMP2

Cited 24 time in Web of Science Cited 23 time in Scopus
Authors
Kim, Yoon Young; Ku, Seung-Yup; Jang, Jiho; Oh, Sun Kyung; Kim, Hee Sun; Kim, Seok Hyun; Choi, Young Min; Moon, Shin Yong
Issue Date
2008-10-31
Publisher
Yonsei University College of Medicine
Citation
Yonsei Med J. 2008;49(5):819-27
Keywords
Bone Morphogenetic Protein 2/*pharmacologyCell Culture Techniques*Cell DifferentiationCell LineCell ProliferationEmbryonic Stem Cells/cytology/*drug effectsHumansMyocytes, Cardiac/*cytologyPluripotent Stem Cells/cytology/drug effectsSignal Transduction
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 (aMHC). 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.
ISSN
0513-5796 (Print)
Language
English
URI
http://hdl.handle.net/10371/67306
DOI
https://doi.org/10.3349/ymj.2008.49.5.819
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College of Medicine/School of Medicine (의과대학/대학원)Obstetrics & Gynecology (산부인과전공)Journal Papers (저널논문_산부인과학전공)
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