Synaptophysin gene expression in neural cells resulted from embryonal carcinoma stem cell differentiation influenced by rat neonate brain extract

Aim: In this study, the effect of newborn rat brain extract to induce neuronal differentiation in P19 embryonal carcinoma (EC) stem cells was investigated.
Material and Newborn rat brain extract was collected in sterile condition and the concentration of its total protein was determined. Then, the amount of cell viability was determined after treatment with brain extract. In order to differentiation, the embryoid bodies resulted from cells suspension culture were exposed to culture medium containing 3% serum that supplemented by the extract, for 7-14 days. Specific staining and real-time PCR methods were applied to evaluate neural differentiation.
Cresyl violet staining confirmed neuronal morphology of the differentiated cells. The gene expression of neural specific was confirmed by real–time PCR. Developing brain extract, which contains numerous neorotrophic factors, could induce expression of synaptophysin (presynaptic membrane protein) and nestin (intermediate filament of stem cells in the neural tube) genes. In addition, the expression of transcription factor Nanog, an important factor for pluripotency and self-renewal of stem cells, decreased under the influence of newborn rat brain extract.
The results of the present study indicated that newborn rat brain extract can induce neuronal phenotype as well as neuronal specific gene expression in P19 stem cells. This study suggests the potential use of combined newborn rat brain extract and stem cell therapy to improvement of deficits in neurodegenerative diseases.