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Introduction: The p19 line of embryonal carcinoma cells develops into neurons, astroglia and fibroblasts after aggregation and exposure to retinoic acid (RA). Dehydroepiandroesteron (DHEA) is a neurosteroid, can increase proliferation of human neural stem cell (NSC) and positively regulated the number of neurons produced. This study was initiated to assess the effect of DHEA on neural progenitor cells derived from p19 embryonal carcinoma stem cells. Methods: p19 cells suspended in DMEM contain 5%FBS into bacterial-greade Petri dishes in the presence RA and DHEA in different concentration for 6 days. Serum concentration decrease to 3% in days 5 and 6. Then collected aggregate and processed for flowcytometry, immunocytochemistery and RT-PCR analysis. Cells were trypsinized for dispersion and replaced in poly L- lysine (10µg/ml) coated tissue culture dishes without RA and DHEA for 4 days. And then difference cells were evaluated by phase contrast microscopy. Results: Flowcytometry analyses of Nestin and Brdu/Nestin showed percent Nestin positive and proliferation Nestin positive cells in different groups DHEA and RA. Brdu/Nestin immunochemistry confirmed proliferation of Nestin positive cells and also RT-PCR analysis show expression of proneural marker and estrogen receptor gens.Result showed that RA +DHEA(1μM) significantly increased the number of Nestin positive and newly formed Nestin positive cells than other groups. Conclusion: Result showed DHEA accompanied RA significantly increased the number of Nestin positive and newly formed Nestin positive cells derived from p19 embryonal carcinoma cells but in comparison to RA cannot induce neural progenitor cells.

Keywords: DHEA, RA, neuron, differentiation, p19 embryonal carcinoma stem cells

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