Modification of a Purification and Expansion Method for Human Embryonic Stem Cell-Derived Cardiomyocytesopen access
- Authors
- Park, Soon-Jung; Bae, Daekyeong; Moon, Sung-Hwan; Chung, Hyung-Min
- Issue Date
- Mar-2013
- Publisher
- KARGER
- Keywords
- Cardiomyocytes; Expansion; Human embryonic stem cells; Purification
- Citation
- CARDIOLOGY, v.124, no.3, pp 139 - 150
- Pages
- 12
- Journal Title
- CARDIOLOGY
- Volume
- 124
- Number
- 3
- Start Page
- 139
- End Page
- 150
- URI
- https://scholarworks.bwise.kr/cau/handle/2019.sw.cau/69858
- DOI
- 10.1159/000346390
- ISSN
- 0008-6312
1421-9751
- Abstract
- Objective:This study aimed to develop a simple and efficient purification method for human embryonic stem cell (hESC)-derived cardiomyocytes (CMs) using a low-glucose culture system. In addition, we investigated whether intercellular adhesion between single hESC-CMs plays a critical role in enhancing proliferation of purified hESC-CMs. Method: hESCs were cultured in suspension to form human embryoid bodies (hEBs) from which similar to 15% contracting clusters were derived after 15-20 days in culture. To purify CMs from contracting hEBs, we first manually isolated contracting clumps that were re-cultured on gelatin-coated plates with media containing a low concentration of glucose. The purified hESC-CMs were cultured at different densities to examine whether cell-cell contact enhances proliferation of hESC-CMs. Results: Purified CMs demonstrated spontaneous contraction and strongly expressed the CM-specific markers cardiac troponin T and slow myosin heavy chain. We investigated the purification efficiency by examining the expression levels of cardiac-related genes and the expression of MitoTracker Red dye. In addition, purified hESC-CMs in low-glucose culture demonstrated a 1.5-fold increase in their proliferative capacity compared to those cultured as single hESC-CMs. Conclusion: A low level of glucose is efficient in purifying hESC-CMs and intercellular adhesion between individual hESC-CMs plays a critical role in enhancing hESC-CM proliferation. Copyright (C) 2013 S. Karger AG, Basel
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