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目的比较两种体外诱导大鼠骨髓间充质干细胞(BMSCs)向γ-氨基丁酸(GABA)能神经元分化的效果。方法从SD大鼠股骨骨髓中分离出BMSCs,进行培养、鉴定。用两种不同的方法进行诱导,方法一设为对照组:碱性成纤维细胞生长因子(bFGF)、表皮生长因子(EGF)、脑源性神经营养因子(BDNF)、全反式维甲酸(ATRA)诱导;方法二设为实验组:ATRA、N5,O2’-二丁酰腺苷3’:5’-环磷酸(Bt2环磷腺苷)(Bt2CAMP)诱导。对分化细胞进行形态学观察和免疫荧光染色。结果两组细胞均出现类似神经细胞的形态变化,实验组细胞变化更明显。两组细胞均能表达nestin和GABA能神经元标记GAD-67,对照组与实验组表达GAD-67的阳性率分别为33.13%±5.12%和41.06%±6.32%,两组相比较具有统计学意义(P<0.05)。结论实验组定向分化为GABA能神经元的效率高于对照组。
Objective To compare the effects of two kinds of BMSCs on the differentiation of γ-aminobutyric acid (GABA) neurons in vitro. Methods BMSCs were isolated from the femur bone marrow of SD rats and cultured. Two different methods were used for induction. The method was as follows: control group: basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), brain-derived neurotrophic factor (BDNF), all- ATRA). The second method was set as the experimental group: ATRA, N5, O2’-dibutyryl adenosine 3 ’: 5’-cyclic phosphatase (Bt2CAMP) induced. Differentiated cells were observed morphologically and immunofluorescently. Results The morphological changes of nerve cells were observed in both groups. The changes of cells in experimental group were more obvious. The positive rates of GAD-67 in nestin and GABAergic neurons were 33.13% ± 5.12% and 41.06% ± 6.32% in control group and experimental group, respectively, which were statistically significant Significance (P <0.05). Conclusion The efficiency of directional differentiation of GABAergic neurons in experimental group is higher than that in control group.