Ps-12: Transplantation of GFP-Expressing Neuron Like Cells Drived from P19 Stem Cells into Midbrain of Chicken Embryo

Bakhshalizadeh SH *, Esmaeili F , Shirzad H , Houshmand F ,


Objective: P19 cells are a line of pluripotent embryonal carcinoma stem cells able to grow continuously in serum-supplemented media. Also p19 cells are amenable to genetic manipulation. when these cells treated with deprenyl can differentiate into cell types similar to those derived from neuroectoderm. The antiparkinsonian effect of deprenyl was reported by several investigators. Because of the limited regenerative potential of adult neural tissue, stem cells may provide a means of surmounting this problem, because they can generate unlimited numbers of cells for use in tissue replacement therapies. Recent reports have shown that many kinds of stem cells can integrate into the chicken embryo and differentiate into various cell types with apparent fusion to the host chicken cells. In this investigation,deprenyl was used to induce neuronal differentiation in GFP-transfected pluripotent P19 cells. Materials and Methods: The cells were cultured using α-MEM medium that supplemented with 15% FBS. The optimal inducing dose of deprenyl was obtained using different concentrations of deprenyl (10-6 – 10-11 M) and the peak response was at 10-8 M. Morphologic and immunofluorescence techniques were used to evaluate the differentiation of the P19 cells, Cresyl violet for the morphologic study, anti-synaptophysin and anti-Beta- Tubulin III antibodies for characterizing the neuronal phenotype of the cells. Then we implanted P19-derived neurons into midbrain of 68-72 hours developed chicken embryo. We used anti-synaptophysin and anti- Beta-Tubulin III and anti-GFP antibodies by using of duble-immunofluorescence techniques in chicken tissue sections to detect the implanted GFP positive P19-derived neurons and follow their migration. Results: The results showed that deprenyl could induce dose-dependent GFP-transfected P19 differentiation into neurons. Also, GFP positive P19-derived neurons could migrate in neuronal system of chicken embryo and make synapse with host tissue cells. Conclusion: According to our finding the combined deprenyl and stem cell therapy can be considered to improve deficits in neurodegenerative diseases.