Circulation Research. 2002;91:501-508
Published online before print August 29, 2002, doi: 10.1161/01.RES.0000035254.80718.91
Published online before print August 29, 2002, doi: 10.1161/01.RES.0000035254.80718.91
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© 2002 American Heart Association, Inc.
Cellular Biology |
Characterization and Enrichment of Cardiomyocytes Derived From Human Embryonic Stem Cells
From the Geron Corporation, Menlo Park, Calif.
Correspondence to Chunhui Xu, PhD, Geron Corporation, 230 Constitution Dr, Menlo Park, CA 94025. E-mail cxu{at}geron.com
Cell replacement therapy is a promising approach for the treatment of cardiac diseases, but is challenged by a limited supply of appropriate cells. We have investigated whether functional cardiomyocytes can be efficiently generated from human embryonic stem (hES) cells. Cardiomyocyte differentiation was evaluated using 3 parent (H1, H7, and H9) hES cell lines and 2 clonal (H9.1 and H9.2) hES cell lines. All cell lines examined differentiated into cardiomyocytes, even after long-term culture (50 passages or 
260 population doublings). Upon differentiation, beating cells were observed after one week in differentiation conditions, increased in numbers with time, and could retain contractility for over 70 days. The beating cells expressed markers characteristic of cardiomyocytes, such as cardiac 
-myosin heavy chain, cardiac troponin I and T, atrial natriuretic factor, and cardiac transcription factors GATA-4, Nkx2.5, and MEF-2. In addition, cardiomyocyte differentiation could be enhanced by treatment of cells with 5-aza-2'-deoxycytidine but not DMSO or retinoic acid. Furthermore, the differentiated cultures could be dissociated and enriched by Percoll density centrifugation to give a population containing 70% cardiomyocytes. The enriched population was proliferative and showed appropriate expression of cardiomyocyte markers. The extended replicative capacity of hES cells and the ability to differentiate and enrich for functional human cardiomyocytes warrant further development of these cells for clinical application in heart diseases.
Key Words: human embryonic stem cells • cardiomyocytes • differentiation • pharmacological responses • cell separation
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