In vitro neural differentiation of bone marrow stromal cells induced by hepatocyte growth factor and glial cell derived neurotrophic factor

摘要

OBJECTIVE: Bone marrow stromal cells (BMSCs) have great potential for cell-based transplantation therapy in treating neurological disease. However, the best combination of various trophic factors to produce full neural differentiation of BMSCs was still unclear. In our study, we aimed to investigate the neural differentiation capacity of rat BMSCs induced by growth factors including hepatocyte growth factor (HGF) and glial cell-derived neurotrophic factor (GDNF). MATERIALS AND METHODS: Cell counting kit-8 (CCK-8) assay, BrdU cell proliferation assay and flow cytometry were implemented to evaluate whether GDNF and HGF had positive effects on the proliferation of BMSCs. Moreover, the expression of neural specific markers in BMSCs was identified using immunofluorescence and quantitative real-time polymerase chain reaction (RT-PCR) at various time points (1, 7, 14 and 21-day post-induction). RESULTS: CCK-8 and BrdU proliferation analyses demonstrated that only HGF treatment had positive effects on the proliferation of BMSCs on the day 14 and 21 after incubation. RT-PCR and immunofluorescence analyses showed that GDNF and HGF elevated the expression of nestin and NCAM, and the combined application of GDNF and HGF has the most significant effect on day 7 after induction. However, at the day of 14 and 21 post-induction, the expression level of nestin and NCAM in GDNF-treatment group was significantly higher than the other three groups. CONCLUSIONS: HGF, not GDNF plays a positive role in BMSCs proliferation, whereas GDNF and HGF are capable of promoting BMSCs to differentiate into neuron-like cells.