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

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)μm,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)×107/L,C组(4.20±0.79)×107/L,差异无统计学意义(P>0.05),但明显高于B组(3.23±0.37)×107/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.

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

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 β-BME and BHA. Group C: a contrast group. The morphology of ADSCs was observed and immunohistology was pedormed. 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.