Scientific Journal of

  Oxytocin efficiency in differentiation of P19c16 stem cells into cardiomyocytes

 Fathi F.¹(PhD), Bagheban Eslami M.R.²(PhD), Ahsan B.¹(MD), Alasvand M.¹(MS),

  Rezaei M.J.¹(PhD), Pirmoradi L.¹(MS), Asahara T.³(MD)

  ¹ School of Medicine, Kordestan University of Medical Sciences

  ² Royan Research Center, Stem Cells Research Group

  ³ Riken Institute, CDB, Japan

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  Abstract

 Background and Objectives

 Recently, P19c16 has been cloned from P19 cells. This stem cell line has mesodermal traits and can be differentiated into cardiomyocytes without embryoid body formation. The present study was designed to investigate the oxytocin effect on differentiation of P19c16 into cardiomyocytes and production of GFP-labeled cardiomyocytes.

 Materials and Methods

 At first, the P19c16 cells were cultured as monolayer and embryoid body then oxytocin with 1 × 10^-7 M concentration was added to culture media as the inducer agent. DMSO was used as an inducer agent in positive control under the same conditions. Differentiated cells underwent morphological and immunocytochemical evaluation. The research in its continuation pursued the transfection of pEGFP-C1 plasmid with P19c16 cells by electroporation method then, stably expressed GFP cells were differentiated into cardiomyocytes by oxytocin.

 Results

 The obtained data indicated that in the experimental group EBs from P19c16 cells could be differentiated into cardiomyocytes whereas monolayer cells could not. In the control group, both EBs and monolayer cells could be differentiated into cardiomyocytes by DMSO. In second part of this study, P19c16 cells were efficiently transfected with GFP, and GFP expressed cells were differentiated into beating cardiomyocytes by oxytocin. The results of morphological and immunocytochemical evaluation confirmed that differentiated cells were cardiomyocytes.

 Conclusions

 Our results showed that the embryoid body formation is necessary for differentiation of P19c16 cells into cardiomyocytes and these cells after stable transfection with GFP could be differentiated into beating cardiomyocytes. It can be concluded that probably P19c16 line is a good stem cell line for cell transplantation in the experimental model of cardiac disease.

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 Key words : Cell differentiation, Cardiomyocytes, Oxytocin, Stem cells , P19