This Article Full Text Full Text (PDF) All Versions of this Article: 107/14/1912    most recent 01.CIR.0000064899.53876.A3v1 Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Takahashi, T. Articles by Lee, R. T. Search for Related Content PubMed PubMed Citation Articles by Takahashi, T. Articles by Lee, R. T. Pubmed/NCBI databases Compound via MeSH Substance via MeSH Hazardous Substances DB ASCORBIC ACID SODIUM ASCORBATE Medline Plus Health Information Stem Cells Related Collections Myogenesis

(Circulation. 2003;107:1912.)
© 2003 American Heart Association, Inc.

Basic Science Reports

Ascorbic Acid Enhances Differentiation of Embryonic Stem Cells Into Cardiac Myocytes

Tomosaburo Takahashi, MD; Bernadette Lord, BS; P. Christian Schulze, MD; Ryan M. Fryer, PhD; Satinder S. Sarang, PhD; Steven R. Gullans, PhD; Richard T. Lee, MD

From the Cardiovascular Division (T.T., P.C.S., B.L., R.T.L.) and Center for Neurological Diseases (R.M.F., S.S.S., S.R.G.), Department of Medicine, Brigham and Women’s Hospital, Harvard Medical School, Boston, Mass.

Correspondence to Richard T. Lee, MD, Partners Research Facility, 65 Landsdowne St, Cambridge, MA 02139. E-mail rlee{at}rics.bwh.harvard.edu

Background— Embryonic stem (ES) cells are capable of self-renewal and differentiation into cellular derivatives of all 3 germ layers. In appropriate culture conditions, ES cells can differentiate into specialized cells, including cardiac myocytes, but the efficiency is typically low and the process is incompletely understood.

Methods and Results— We evaluated a chemical library for its potential to induce cardiac differentiation of ES cells in the absence of embryoid body formation. Using ES cells stably transfected with cardiac-specific -cardiac myosin heavy chain (MHC) promoter-driven enhanced green fluorescent protein (EGFP), 880 compounds approved for human use were screened for their ability to induce cardiac differentiation. Treatment with ascorbic acid, also known as vitamin C, markedly increased the number of EGFP-positive cells, which displayed spontaneous and rhythmic contractile activity and stained positively for sarcomeric myosin and -actinin. Furthermore, ascorbic acid induced the expression of cardiac genes, including GATA4, -MHC, and ß-MHC in untransfected ES cells in a developmentally controlled manner. This effect of ascorbic acid on cardiac differentiation was not mimicked by the other antioxidants such as N-acetylcysteine, Tiron, or vitamin E.

Conclusions— Ascorbic acid induces cardiac differentiation in ES cells. This study demonstrates the potential for chemically modifying the cardiac differentiation program of ES cells.

Key Words: stem cells • ascorbic acid • myocytes

This article has been cited by other articles:

				S. Takei, H. Ichikawa, K. Johkura, A. Mogi, H. No, S. Yoshie, D. Tomotsune, and K. Sasaki Bone morphogenetic protein-4 promotes induction of cardiomyocytes from human embryonic stem cells in serum-based embryoid body development Am J Physiol Heart Circ Physiol, June 1, 2009; 296(6): H1793 - H1803. [Abstract] [Full Text] [PDF]

				T. Tokunou, R. Miller, P. Patwari, M. E. Davis, V. F. M. Segers, A. J. Grodzinsky, and R. T. Lee Engineering insulin-like growth factor-1 for local delivery FASEB J, June 1, 2008; 22(6): 1886 - 1893. [Abstract] [Full Text] [PDF]

				H. Sadek, B. Hannack, E. Choe, J. Wang, S. Latif, M. G. Garry, D. J. Garry, J. Longgood, D. E. Frantz, E. N. Olson, et al. Cardiogenic small molecules that enhance myocardial repair by stem cells PNAS, April 22, 2008; 105(16): 6063 - 6068. [Abstract] [Full Text] [PDF]

				A. Raya, I. Rodriguez-Piza, B. Aran, A. Consiglio, P.N. Barri, A. Veiga, and J.C. I. Belmonte Generation of Cardiomyocytes from New Human Embryonic Stem Cell Lines Derived from Poor-quality Blastocysts Cold Spring Harb Symp Quant Biol, January 1, 2008; 73(0): 127 - 135. [Abstract] [PDF]

				R. Kitamura, T. Takahashi, N. Nakajima, K. Isodono, S. Asada, H. Ueno, T. Ueyama, T. Yoshikawa, H. Matsubara, and H. Oh Stage-Specific Role of Endogenous Smad2 Activation in Cardiomyogenesis of Embryonic Stem Cells Circ. Res., July 6, 2007; 101(1): 78 - 87. [Abstract] [Full Text] [PDF]

				M. Hosseinkhani, H. Hosseinkhani, A. Khademhosseini, F. Bolland, H. Kobayashi, and S. P. Gonzalez Bone Morphogenetic Protein-4 Enhances Cardiomyocyte Differentiation of Cynomolgus Monkey ESCs in Knockout Serum Replacement Medium Stem Cells, March 1, 2007; 25(3): 571 - 580. [Abstract] [Full Text] [PDF]

				A. M. Singh, F.-Q. Li, T. Hamazaki, H. Kasahara, K.-I. Takemaru, and N. Terada Chibby, an Antagonist of the Wnt/{beta}-Catenin Pathway, Facilitates Cardiomyocyte Differentiation of Murine Embryonic Stem Cells Circulation, February 6, 2007; 115(5): 617 - 626. [Abstract] [Full Text] [PDF]

				R. Zhou, P. D. Acton, and V. A. Ferrari Imaging Stem Cells Implanted in Infarcted Myocardium J. Am. Coll. Cardiol., November 21, 2006; 48(10): 2094 - 2106. [Abstract] [Full Text] [PDF]

				X.-M. Guo, Y.-S. Zhao, H.-X. Chang, C.-Y. Wang, L.-L. E, X.-A. Zhang, C.-M. Duan, L.-Z. Dong, H. Jiang, J. Li, et al. Creation of Engineered Cardiac Tissue In Vitro From Mouse Embryonic Stem Cells Circulation, May 9, 2006; 113(18): 2229 - 2237. [Abstract] [Full Text] [PDF]

				K. Fukuda and S. Yuasa Stem Cells as a Source of Regenerative Cardiomyocytes Circ. Res., April 28, 2006; 98(8): 1002 - 1013. [Abstract] [Full Text] [PDF]

				S. P. Raikwar, T. Mueller, and N. Zavazava Strategies for Developing Therapeutic Application of Human Embryonic Stem Cells Physiology, February 1, 2006; 21(1): 19 - 28. [Abstract] [Full Text] [PDF]

				D. Hakuno, T. Takahashi, J. Lammerding, and R. T. Lee Focal Adhesion Kinase Signaling Regulates Cardiogenesis of Embryonic Stem Cells J. Biol. Chem., November 25, 2005; 280(47): 39534 - 39544. [Abstract] [Full Text] [PDF]

				R. Passier, D. W.-v. Oostwaard, J. Snapper, J. Kloots, R. J. Hassink, E. Kuijk, B. Roelen, A. B. de la Riviere, and C. Mummery Increased Cardiomyocyte Differentiation from Human Embryonic Stem Cells in Serum-Free Cultures Stem Cells, June 1, 2005; 23(6): 772 - 780. [Abstract] [Full Text] [PDF]

				G. Keller Embryonic stem cell differentiation: emergence of a new era in biology and medicine Genes & Dev., May 15, 2005; 19(10): 1129 - 1155. [Abstract] [Full Text] [PDF]

				E. S. Ng, L. Azzola, K. Sourris, L. Robb, E. G. Stanley, and A. G. Elefanty The primitive streak gene Mixl1 is required for efficient haematopoiesis and BMP4-induced ventral mesoderm patterning in differentiating ES cells Development, March 1, 2005; 132(5): 873 - 884. [Abstract] [Full Text] [PDF]

				M. E. Davis, J.P. M. Motion, D. A. Narmoneva, T. Takahashi, D. Hakuno, R. D. Kamm, S. Zhang, and R. T. Lee Injectable Self-Assembling Peptide Nanofibers Create Intramyocardial Microenvironments for Endothelial Cells Circulation, February 1, 2005; 111(4): 442 - 450. [Abstract] [Full Text] [PDF]

				S. Kanno, P. K. M. Kim, K. Sallam, J. Lei, T. R. Billiar, and L. L. Shears II Nitric oxide facilitates cardiomyogenesis in mouse embryonic stem cells PNAS, August 17, 2004; 101(33): 12277 - 12281. [Abstract] [Full Text] [PDF]