Investigation of Neuronal Phenotype Injection in P19 Stem Cells Under the Influence of Rat Infant Extract and Deprenil

Parkinson's disease cause by selective loss of a group of dopaminergic cells in the Substantial nigra. One of the treatments for this disease is the displacement of lost cells by stem cells. The purpose of this study was to investigate the effect of inducers of extract of neonatal rat brain and deprenyl on neural differentiation of P19 embryonic carcinoma cells. P19 stem cells were cultured in alpha-MEM medium supplemented with 10% FBS serum. Then, the cells were cultured on low-adhesion culture plates to create pseudo-embryonic bodies. In order to differentiate, germ cells were cultured in medium containing 3% FBS serum and extract of neonatal rat brain, deprenyl and mixture of both extract for 28 days. Immunofluorescence assay and Real-time PCR was applied to detect and evaluate the expression specific proteins of neurons including synaptophysin, beta-tubulin 3 and nestin, respectively. Regarding to immunofluorescence results, differentiated P19 cells, positively affected by three inducers such as nestin, synaptophysin and beta-3. Relative expression of transcription factors of nestin, synaptophysin in cells differentiated by all three inducer using Real-time PCR demonstrated that in P19 embryonic carcinoma cells, Nestin and Synaptophysin expression increased in the second week, while, thier expression significantly reduced in the third and fourth weeks. The results of this study indicated P19 embryonic carcinoma stem cells are potential to differentiate into neurons, especially dopaminergic phenotype, following laboratory conditions and inducing factors