An in vitro system that supports primordial germ cells (PGCs) survival and proliferation is useful for enhancement of these cells and efficient transplantation in infertility disorders. One approach is cultivation of PGCs under proper conditions that allow self-renewal and proliferation of PGCs. For this purpose, we compared the effects of different concentrations of retinoic acid (RA), and the effect of PGCs co-culture with STO cells on the proliferation of embryonic stem cells (ESCs)- derived PGCs. One-day-old embryoid body (EB) was cultured for 4 days in simple culture system at the presence of 5 ng/ml BMP4 (SCB group) for PGC induction. For PGC enrichment, ESCs derived germ cells cultured for 7 days in the presence of different doses (0-5 µM) of RA both in the simple and STO co-culture systems. At the end of the culture period, viability and proliferation rates were assessed and expression of Mvh, α6 integrin, β1 integrin, Stra8 and Piwil2 was evaluated using quantitative PCR. Also, the inductive effects were investigated immunocytochemically with Mvh and CDH1 on the selected groups. Immunocytochemistry/PCR results showed higher expression of Mvh, the PGC-specific marker, in 3 µM RA concentrations on the top of the STO feeder layer. Meanwhile, assessment of the Stra8 mRNA and CDH1 protein, the specific makers for spermatogonia, showed no significant differences between groups. Based on the results, it seems that in the presence of 3 µM RA on top of the STO feeder layer cells, the majority of the cells transdifferentiated into germ cells were PGC.
- Embryonic stem cell
- retinoic acid
- primordial germ cell
- ©2016 The Author(s)
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