许旺细胞诱导下脂肪来源干细胞向神经细胞方向分化的研究

Schwann cells induced neuronal differentiation of adipose-derived stem cells （ADSCs）

目的探讨脂肪来源干细胞（ADSCs）和SD大鼠许旺细胞（Schwann cells）利用Transwell非接触式共培养时生长增殖特性和向神经元样细胞分化的规律。方法取1月龄SD大鼠腹股沟处脂肪组织。按酶原消化法获取脂肪来源干细胞，同时取大鼠坐骨神经按改良植块法获取许旺细胞。取P3代脂肪来源干细胞分三组：A组使用ADSCs和许旺细胞构建Transwell非接触式共培养体系，B组利用β-巯基乙醇（BME）、丁酸酯羟基茴香醚（BHA）作为脂肪来源干细胞诱导因子使其向神经方向诱导，C组脂肪干细胞为对照组。比较各组细胞形态，观察脂肪来源干细胞代谢、轴突生长、免疫荧光染色情况，并做统计学分析。结果A组脂肪来源干细胞部分分化为具有细长突起的细胞，B组诱导培养后，大多数存活的细胞也同样转变为类似于神经元的形态，A组突起长度（11．64±1．64）μm，B组突起长度（13．03±1．35）±m，差异有统计学意义（P〉0．05），细胞生长代谢旺盛。A组B组诱导分化后的细胞NF表达阳性率分别为（54．16±9．35）％和（62．25±8．95）％，两者差异无统计学意义，而GFAP染色阴性。在细胞数量上，A组为（4．08±0．68）×10^7/L，C组（4．20±0．79）×10^7／L，差异无统计学意义（P〉0．05），但明显高于B组（3．23±0．37）×10^7／L（P〈0．05）。结论许旺细胞能促进脂肪来源干细胞向神经方向分化，效果与使用β—BME加BHA试剂诱导的结果相近，并能有效防止脂肪来源干细胞损伤。

Objective To investigate the function of SD rats Schwann cells on the growth and differentiation of adipose-derived stem cells（ADSCs） when they are co-cultured in Transwell culture system. Methods The ADSCs harvested from SD rats were divided into 3 groups. Group A： ADSCs and Schwann cells were cultured in Transwell indirect co-culture system. Group B： ADSCs were induced by [3-BME and BHA. Group C： a contrast group. The morphology of ADSCs was observed and immunohistology was performed. Results ADSCs of group A and group B were partly differentiated to the cells which had long neurite,and had a positive result in the NF staining, but negative in the GFAP staining. Group A shows no great difference in neurite length compared with group B（P 〈 0.05）, and no great difference in cell quantity compared with group C （P 〈 0.05）. Conclusion Schwann cells of SD rat have a growth supportive and neuronal differentiation inducing effect on ADSCs.