Document Type : Original Article
Stem Cells and Developmental Biology Department, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran
HIV & Hepatitis Research Center, Shiraz University of Medical Science, Shiraz, Iran
Developmental Biology Department, University of Science and Culture, Tehran, Iran
Objective: Dehydroepiandroesteron (DHEA) is a neurosteroid with potential effect on neurogenesis, neuronal survival and proliferation of neural progenitor cells. However there is no direct evidence for its biological effect during the differentiation of stem cell-derived neurons. The p19 line of embryonal carcinoma cells develops into neurons, astroglia and fibroblasts after exposure to retinoic acid (RA). This study was initiated to assess the effect of DHEA on neural cells derived from p19 embryonal carcinoma stem cells.
Materials and Methods: P19 cells were suspended in dulbecco’s modified eagle’s medium (DMED) containing fetal bovine serum (FBS) in bacterial-grade petri dishes in the presence of RA, DHEA and RA+DHEA for 6 days. Then cells were trypsinized for dispersion and replaced in poly L- lysine (10μg/ml) coated tissue culture dishes without RA and DHEA for 4 days. The expression of neural markers Map-2, Tau, beta-tubulin III- clone Juj (Tuj1), astrocyte marker GFAP and the percent of neurotransmitters tyrosin hydroxylase, glutamate, serotonin and actyl cholin transferase were evaluated by flowcytometry, immunocytochemistry and RT-PCR analysis.
Results: Flowcytometry analysis showed that about 63 ± 3% of the cells express neuronal marker Tuj1 and about 5 ± 1% of the cells express tyrosine hydroxylase neurotransmitters in RA treated groups. However when RA and DHEA were added to the culture medium, Tuj1 expression increased to about 74 ± 1% and tyrosine hydroxylase expression increased to 23 ± 2%. Conclusion: Results showed that DHEA accompanied RA increased the number of Tuj1 and dopaminergic neurons that were derived from p19 embryonal carcinoma stem cells.