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目的探讨碱性成纤维细胞生长因子(Basic fibroblast growth factor,bFGF)体外作用于骨髓间充质干细胞(Mesen-chymal stem cells,MSCs)后,诱导其向神经元样细胞和多巴胺能神经元样细胞定向分化的情况。方法从鼠骨髓中获得MSCs,培养传代,用MTT法检测bFGF对骨髓MSCs生长的影响。10 ng/ml bFGF作用2 d后,通过IBMX、细胞因子bFGF、GDNFI、L-1β、中脑神经胶质细胞条件培养基和中脑神经细胞膜碎片等分组联合诱导骨髓MSCs向神经元样细胞、多巴胺能神经元样细胞分化,免疫细胞化学方法鉴定特异标志NSE、MAP-2a,b和TH的表达。结果在一定范围内,bFGF对骨髓MSCs具有明显的促增殖作用。分化的神经元样细胞表达NSE、MAP-2a,b和TH,联合应用GDNFI、L-1β、中脑条件培养基和中脑神经细胞膜碎片诱导7 d后,NSE阳性率为(27.774±2.747)%,MAP-2a,b为(28.006±3.080)%,TH为(3.098±0.352)%。结论体外骨髓MSCs被诱导分化成神经元样细胞和多巴胺能神经元样细胞,为帕金森等中枢神经系统疾病的细胞移植治疗奠定了基础。
Objective To investigate the effect of basic fibroblast growth factor (bFGF) on the proliferation of neuron-like cells and dopaminergic neuron-like cells (MSCs) after they were treated with mesenchymal stem cells (MSCs) Directional differentiation of the situation. Methods MSCs were obtained from rat bone marrow and cultured for passage. The effects of bFGF on the growth of bone marrow MSCs were detected by MTT assay. After being treated with 10 ng / ml bFGF for 2 days, MSCs were induced to differentiate into neuron-like cells by IBMX, cytokine bFGF, GDNFI, L-1β, mesencephalic glial cell conditioned media and mesangial nerve cell membrane fragments, Dopaminergic neuron-like cell differentiation and immunocytochemistry were used to identify the expression of specific markers NSE, MAP-2a, b and TH. Results In a certain range, bFGF had a significant effect on proliferation of bone marrow MSCs. Differentiated neuron-like cells expressed NSE, MAP-2a, b and TH, and the positive rate of NSE was (27.774 ± 2.747) after 7 days of induction by GDNFI, L-1β, midbrain conditioned medium and midbrain nerve membrane fragments, %, MAP-2a, b (28.006 ± 3.080)%, TH (3.098 ± 0.352)%. Conclusion In vitro bone marrow MSCs are differentiated into neuron-like cells and dopaminergic neuron-like cells, which lays a foundation for the treatment of central nervous system diseases such as Parkinson’s disease.