Cardiac differentiation of P19CL6 cells by oxytocin

Fathi, Fardin; Murasawa, Satoshi; Hasegawa, Satoshi; Asahara, Takayuki; Kermani, Abbas Jafari; Mowla, Seyed Javad

doi:10.1016/j.ijcard.2008.01.046

« Previous Next »International Journal of Cardiology
Volume 134, Issue 1 , Pages 75-81, 1 May 2009

Cardiac differentiation of P19CL6 cells by oxytocin

Fardin Fathi
- KDRC, Faculty of Medicine, Kurdistan University of Medical Sciences, Sanandaj, Iran
- Corresponding author. Tel.: +98 871 6661830; fax: +98 871 6664674.
Satoshi Murasawa
- Group of Vascular Regeneration Research, Kobe Institute of Biomedical Research and Innovation/Riken Center for Developmental Biology, 2-2 Minatojima-Minamimachi, Chuo-ku, Kobe 650-0047, Japan
Satoshi Hasegawa
- Group of Vascular Regeneration Research, Kobe Institute of Biomedical Research and Innovation/Riken Center for Developmental Biology, 2-2 Minatojima-Minamimachi, Chuo-ku, Kobe 650-0047, Japan
Takayuki Asahara
- Group of Vascular Regeneration Research, Kobe Institute of Biomedical Research and Innovation/Riken Center for Developmental Biology, 2-2 Minatojima-Minamimachi, Chuo-ku, Kobe 650-0047, Japan
Abbas Jafari Kermani
- Department of Genetics, Faculty of Basic Sciences, Tarbiat Modares University, Tehran, Iran
Seyed Javad Mowla
- Department of Genetics, Faculty of Basic Sciences, Tarbiat Modares University, Tehran, Iran

Received 9 May 2007; received in revised form 31 August 2007; accepted 14 January 2008. published online 06 June 2008.

Abstract

Background

It has been reported that P19 embryonal carcinoma (EC) cells differentiate into beating cardiomyocytes under the action of oxytocin (OT). It has been suggested that dimethylsulfoxide (DMSO) acts via the oxytocin/oxytocin receptor pathway because an oxytocin receptor antagonist not only blocks oxytocin-induced cardiomyocyte differentiation, but also blocks DMSO-induced differentiation. In this study, the differentiation ability of OT was tested using P19CL6 cells.

Methods

P19CL6 cells were cultured as a confluent monolayer and aggregated cells. OT was then added to culture media as an inducing agent. The cells treated with 1% DMSO were used as a positive control group. Differentiated cells were evaluated morphologically and immunocytochemically, as well as by RT-PCR. In addition, a stable line of green fluorescent protein (GFP)-expressing P19CL6 cells were differentiated into beating cardiomyocytes by OT.

Results

Aggregated P19CL6 cells could be differentiated into cardiomyocytes, whereas monolayer cells could not differentiate and express specific cardiac muscle marker genes. In the control group, both aggregates and monolayer cells could be differentiated into cardiomyocytes by DMSO. In addition, GFP-expressing P19CL6 cells differentiated efficiently into beating cardiomyocytes when treated with OT. The results of all evaluations confirmed that the differentiated cells were cardiomyocytes.

Conclusions

We concluded that embryoid body formation (cell aggregation) is necessary for the differentiation of P19CL6 cells into cardiomyocytes when using OT as an inducer agent. Furthermore, because of the high rate of differentiation efficiency, GFP-expressing cardiomyocytes derived from P19CL6 cells have the potential to be used for regenerative therapies in experimental models.

Keywords: Cardiomyocytes, Cell differentiation, Embryoid body formation, Oxytocin, P19CL6 cells