Development of a novel two-dimensional directed differentiation system for generation of cardiomyocytes from human pluripotent stem cellsopen access
- Authors
- Moon, Sung-Hwan; Ban, Kiwon; Kim, Changhoon; Kim, Sang-Sung; Byun, Jaemin; Song, Ming-Ke; Park, In-Hyun; Yu, Shan Ping; Yoon, Young-Sup
- Issue Date
- Sep-2013
- Publisher
- ELSEVIER IRELAND LTD
- Keywords
- Human embryonic stem cells; Human induced pluripotent stem cells; Cardiomyocytes; Directed differentiation
- Citation
- INTERNATIONAL JOURNAL OF CARDIOLOGY, v.168, no.1, pp 41 - 52
- Pages
- 12
- Journal Title
- INTERNATIONAL JOURNAL OF CARDIOLOGY
- Volume
- 168
- Number
- 1
- Start Page
- 41
- End Page
- 52
- URI
- https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/69852
- DOI
- 10.1016/j.ijcard.2012.09.077
- ISSN
- 0167-5273
1874-1754
- Abstract
- 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 similar to 90% showed CM phenotypes confirmed by an a-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. (c) 2012 Elsevier Ireland Ltd. All rights reserved.
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