摘要
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.
出处
《中华显微外科杂志》
CSCD
北大核心
2009年第2期119-122,I0005,共5页
Chinese Journal of Microsurgery
基金
2006复旦青年骨干、上海市卫生局科研基金(2007041)
973课题基金赞助(2005CB522604)
关键词
许旺细胞
脂肪干细胞
神经分化
Schwann cell
Adipose-derived stem cells
Neural differentiation