体外诱导大鼠骨髓间充质干细胞向GABA能神经元分化

目的研究骨髓间充质干细胞（BMSCs）在体外向γ-氨基丁酸（GABA）能神经元方向的分化。方法大鼠股骨骨髓分离出BMSCs,培养传代,通过10%胎牛血清（FBS）、20 ng/ml表皮生长因子（EGF）和改良Eagle培养基（DMEM/F12）预诱导24 h,随之加入含10 ng/ml碱性成纤维细胞生长因子（bFGF）、10 ng/ml骨形成蛋白-2（BMP-2）和10 ng/ml脑源性神经营养因子（BDNF）作用48 h后,再以10μmol/L全反式维甲酸（ATRA）培养48 h,最后用40 mmol/L KCl刺激15 min完成诱导分化。对照组用10%FBS和DMEM/F12培养不添加任何诱导因子。形态学观察和Western blot对分化后的细胞进行鉴定分析。结果实验组细胞在形态学上表现出典型的神经元样细胞形态,对照组无明显变化;Western blot分析知实验组Ⅰ和实验组Ⅱ都有分化为GABA能神经元的趋势,但实验组Ⅱ的分化率高于实验组Ⅰ。结论BMSCs可在体外分化为GABA能神经元。

Rat bone marrow mesenchymal stem cells differentiated into GABAergic neurons in vitro

Objective To study the bone marrow mesenchymal stem cells （BMSCs） to differentiate into gamma-aminobutyric acid （GABAergic） neurons in vitro. Methods BMSCs were isolated from bone marrow of rat and cultured and transferred by using 10% fetal bovine serum （FBS）, 20 ng/ml epidermal growth factor （EGF） and Dulbecco minimum essential medium/F12 （ DMEM/F12） to induce BMSCs for 24 h in advance. After adding 10 ng/ml basic fibroblast growth factor （bFGF）, 10 ng/ml bone morphogenetic protein-2 （BMP-2） and 10 ng/ml brain-derived neurotrophic factor （BDNF） for 48 h, cells were cultivated with 10 μmol/L all transretinoic acid （ATRA） for 48 h. Correspondingly 40 mmoL/L KCl was used to stimulate for 15 min to complete BMSCs differentiation. The control group did not add any induced factor except 10% FBS and DMEM / F12. The morphology observation and Western blot were used to carry on the appraisal analysis of differentiated cells. Results The cell morphology of experimental group manifested typically neuronal morphology, but little change was observed in the control group. Western blot analysis showed that experimental group Ⅰ and experimental group Ⅱ had the tendency to differentiate into GABAergic neurons, and the differentiation rate of experimental group Ⅱ was higher than that of experimental group Ⅰ. Conclusion BMSCs can differentiate into GABAergic neurons in vitro.