Effect Of Grape Seed Proanthocyanidins On The Apoptosis Of Neuron-like Cells Differentiated From Mesenchymal Stem Cells

Objective To study the effect of grape seed proanthocyanidins on the apoptosis of neuron-like cells differentiated from mesenchymal stem cells in vitro and the optimal dose.Methods BMSCs were isolated primarily from bone marrow of adult WISTAR rats and purified with adherence cultivation. The different dosis (20mg/L,40mg/L,80mg/L) ofGSP interfere for 24 h which are 5th generations. Take the blank as control group. Then we use lmmol/L B-mercaptoethanol (BME) containing 20% fetal bovine serum (FBS) to preinduce BMSCs for 24 h. Serum-free medium that contained 40 ng/mL basic fibroblast growth factor (bFGF) are used as induction liquid for another 24 h. The expression of neuronspecificenolase (NSE) and microtubule-associated protein 2(MAP-2) are detected by using immunocytochemistry method as specific neuron marker. To test the vigor of neuron-like cells by using MTT method. To test the fluorescent probe labeled intracellular free calcium concentration by flow cytometry. To test the interclass apoptosis rate by Annexin V apoptosis kit.Results The BMSCs of WISTAR rats were successfully cultivated. All protocols induced BMSCs to differentiate into neuron-like cells which expressed NSE and MAP-2. The GSP interfered groups get better condition, vigor and living cell proportion. Conclusion GSP can improve the vigor, antagonize free radicle and decrease apoptosis of neuron-like cells differentiated from BMSCs.