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In vitro differentiation of embryonic stem cells into cardiomyocytes or skeletal muscle cells is specifically modulated by retinoic acid

Abstract

Pluripotent embryonic stem cells (ES cells) differentiating via embryo-like aggregates (embryoid bodies) into derivatives of the primary germ layers were used as a model system to investigate the time- and concentration dependent effects of retinoic acid (RA) on the in vitro differentiation pattern. When ES cells, cultivated normally under conditions resulting in cardiomyocyte differentiation, were treated during the first 2 days of embryoid body formation with high RA concentrations (10−9 to 10−7 M) a strong inhibition of cardiogenesis was found. ES cells differentiating as embryoid bodies and treated with the same RA concentration between the 5th and 7th day resulted in a slight induction of cardiogenesis. In contrast, incubation of embryoid bodies with 10−8 and 10−7 M RA between the 2nd and 5th day of embryoid body development resulted in a total inhibition of cardiogenesis but in an induction of myogenesis. This was demonstrated by indirect immunofluorescence and, as shown by reverse transcription- polymerase chain reaction (RT-PCR), by the time- and concentration-dependent inhibition of transcription of cardiac-specific α- and β-cardiac myosin heavy chain (MHC) genes, and the induction of transcription of skeletal muscle-specific myogenin. In addition, using the whole-cell patch-clamp technique, these skeletal myocytes were functionally characterized by the expression of tissue-specific Ca2+ channels and nicotinic cholinoceptors. In summary, a specific effect of RA on ES cell differentiation in the embryoid body resulting in a switch from cardiogenesis to myogenesis and an induction of neuronal cells was found.

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Wobus, A.M., Rohwedel, J., Maltsev, V. et al. In vitro differentiation of embryonic stem cells into cardiomyocytes or skeletal muscle cells is specifically modulated by retinoic acid. Roux's Arch Dev Biol 204, 36–45 (1994). https://doi.org/10.1007/BF00189066

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  • DOI: https://doi.org/10.1007/BF00189066

Key words

  • Mouse embryonic stem cells
  • Differentiation
  • Cardiomyocytes
  • Skeletal muscle cells
  • Retinoic acid