Document Detail

Development of a novel two-dimensional directed differentiation system for generation of cardiomyocytes from human pluripotent stem cells.
MedLine Citation:
PMID:  23044428     Owner:  NLM     Status:  MEDLINE    
BACKGROUND: Human pluripotent stem cells (hPSCs) hold great promise for treating ischemic heart disease. However, current protocols for differentiating hPSCs either result in low yields or require expensive cytokines.
METHODS: Here we developed a novel two dimensional (2D) stepwise differentiation system that generates a high yield of cardiomyocytes (CMs) from hPSCs without using special cytokines. Initially, undifferentiated hPSCs were transferred onto Matrigel-coated plates without forming embryoid bodies (EBs) for a few days and were cultured in bFGF-depleted human embryonic stem cells (hESCs) medium. When linear cell aggregation appeared in the margins of the hPSC colonies, the medium was changed to DMEM supplemented with 10% fetal bovine serum (FBS). Thereafter when cell clusters became visible, the medium was changed to DMEM with 20% FBS.
RESULTS AND CONCLUSIONS: At about two weeks of culture, contracting clusters began to appear and the number of contracting clusters continuously increased, reaching approximately 70% of all clusters. These clusters were dissociated by two-step enzyme treatment to monolayered CMs, of which ~90% showed CM phenotypes confirmed by an α-myosin heavy chain reporter system. Electrophysiologic studies demonstrated that the hPSC-derived CMs showed three major CM action potential types with 61 to 78% having a ventricular-CM phenotype. This differentiation system showed a clear spatiotemporal role of the surrounding endodermal cells for differentiation of mesodermal cell clusters into CMs. In conclusion, this system provides a novel platform to generate CMs from hPSCs at high yield without using cytokines and to study the development of hPSCs into CMs.
Sung-Hwan Moon; Kiwon Ban; Changhoon Kim; Sang-Sung Kim; Jaemin Byun; Ming-Ke Song; In-Hyun Park; Shan Ping Yu; Young-Sup Yoon
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.     Date:  2012-10-06
Journal Detail:
Title:  International journal of cardiology     Volume:  168     ISSN:  1874-1754     ISO Abbreviation:  Int. J. Cardiol.     Publication Date:  2013 Sep 
Date Detail:
Created Date:  2013-09-09     Completed Date:  2014-05-06     Revised Date:  2014-09-19    
Medline Journal Info:
Nlm Unique ID:  8200291     Medline TA:  Int J Cardiol     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  41-52     Citation Subset:  IM    
Copyright Information:
Copyright © 2012 Elsevier Ireland Ltd. All rights reserved.
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MeSH Terms
Cell Differentiation / physiology*
Induced Pluripotent Stem Cells / physiology*,  transplantation*
Myocardial Infarction / pathology,  surgery
Myocytes, Cardiac / physiology*,  transplantation*
Pluripotent Stem Cells / physiology,  transplantation
Primary Cell Culture / methods*,  trends
Rats, Nude
Grant Support

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine

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