胶质细胞源性神经营养因子诱导骨髓基质细胞向多巴胺能神经元分化的观察

Observation of Differentiation of Bone Marrow Stromal Cells into Dopaminergic Neuron Induced by Glia-derived Neurotrophic Factor

目的探讨胶质细胞源性神经营养因子(GDNF)在体外能否诱导骨髓基质细胞(BMSCs)向多巴胺(DA)能神经元分化及可能机制。方法无菌条件下,抽取成年SD大鼠胫骨内骨髓组织,分离制备成单细胞悬液进行培养。将增殖传代至第5代的BMSCs随机分为GDNF诱导组和对照组。继续培养7 d后,应用BrdU/GFAP、BrdU/NeuN和TH免疫荧光单标和双标技术检测BMSCs增殖和分化情况。结果两组BMSCs继续培养7 d后,增殖仍然活跃,有部分细胞向神经元和胶质样细胞分化,呈BrdU/GFAP、BrdU/NeuN和TH阳性表达,但GDNF组的增殖力更强,向神经元和TH神经元分化的数量明显多于对照组(P<0.05)。结论GDNF能促进BMSCs的增殖和诱导BMSCs分化成神经元和胶质样细胞,其中少部分可分化为TH神经元(即DA能神经元)。

Objective To observe to differentiation of bone marrow stromal cells （BMSCs） derived from adults rats into dopaminergic neurons in vitro induced by glia-derived neurotrophic factor（GDNF）. Methods BMSCs were isolated from the bone narrow of tibia cavity of adult SD rats and cultured. BMSCs used in the study were passage 5 cells which were randomly divided into GDNF induction group and control group. The proliferation and differentiation of BMSCs were identified by using immnohistochemical and immunofluorescence techniques. Results BMSCs showed significant proliferation and differentiation 7 days after the culture in both the groups. A part of BMSCs differentiated into the neuron-like cells which expressed Brdu/NeuN and glia-like cells which expressed Brdu/GFAP. A few of the BMSCs differentiated into TH-positive neurons （i.e. dopaminergic neurons）. The numbers of Brdu/NeuN and TH-positive cells in GDNF induction group were significantly more than those in the control group （P〈0.05）. Conclusions The BMSCs can proliferate and differentiate into neuron-like cells and TH-positive neurons by the induction of GDNF.