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Biochem Biophys Res Commun. 2011 Dec 9;416(1-2):51-7. doi: 10.1016/j.bbrc.2011.10.140. Epub 2011 Nov 6.

High density cultures of embryoid bodies enhanced cardiac differentiation of murine embryonic stem cells.

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Yale Cardiovascular Research Center, Department of Internal Medicine, Yale Stem Cell Center, Yale School of Medicine, New Haven, CT 06520, USA.


Murine embryonic stem cell (mESC)-derived cardiomyocytes represent a promising source of cells for use in the development of models for studying early cardiac development as well as cell-based therapies in postnatal pathologies. Here, we report a highly efficient cardiac differentiation system in which high density embryoid body (EB) cultures leads to a marked increase of cardiomyocytes production from multiple mESC lines without the addition of any cardiogenic growth factors. Our results show that high density EB cultures significantly increase the yield of functional cardiomyocytes, which express typical cardiac markers, exhibit normal rhythmic Ca(2+) transients, and respond to both β-adrenergic and electric stimulations. During the differentiation period, the inhibition of bone morphogenetic protein (BMP) signaling significantly attenuates the increase of cardiac differentiation as well as the increased expression of cardiac-specific genes, NK2 transcription factor related 5 (Nkx2.5) and myosin light chain 2v (Mlc2v) by high density EB cultures. Therefore, we believe that we offer a novel and efficient means of cardiomyocyte production for practical use of mESCs in cardiac regenerative medicine.

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