Assessment of neurotrophic factors expression and cell proliferation in the coculture of neural and mesenchymal stem cells

Abstract

Background and Objective: Neurotrophic factors are diffusible polypeptides that have critical roles in survival, proliferation and differentiation of stem cells. This study was done to assess the role of neurotrophic factors (CNTF‎, BDNF, ‎GDN‎F, ‎NT-‎3‎) expression and proliferation rate of neural stem cells (NSCs) in coculture with mesenchymal stem cells (MSCs). Materials and Methods: In this experimental study, NSCs and MSCs were isolated from adult Wistar rat. Initially, NSCs was harvested from temporal lobe after mechanical digestion by a sterile flamed Pasteur pipette and enzymatic digestion with trypsin and Dnase. The cell suspension was cultivated in a flask with DMEM/F12 medium supplemented with 10% FBS 100U/ml Penicillin and 100 mg/ml Streptomycin. To obtain MSCs, bone marrow of femur and tibia bones were flashed out and cultured. MSCs and NSCs‎ cocultured by transwell ‎system in DMEM/F12 medium supplemented with 10% FBS 100U/ml Penicillin and 100 mg/ml Streptomycin. Haemocytometer, immunocytochemistry and RT-PCR methods were performed to identify and evaluate cell proliferation, purity levels and neurotrophic factors expression. Results: There ‎is‎ no differences in NTFs profile of ‎neurotrophic‎ factors expression between ‎coculture ‎group‎ ‎and‎ control ‎NSCs, but interactions between MSCs and NSCs significantly promoted NSCs proliferation (P<0.05). Conclusion: This study showed that coculture of NSCs with MSCs might be prfered in cell-therapy than‎ NSCs.‎

Keywords

• Stem cell
• Neural cell
• Mesenchymal cell
• Coculture
• CNTF‎
• BDNF
• ‎GDN‎F
• ‎NT-‎3