脂肪来源干细胞诱导分化为神经元样细胞的实验研究

Adipose-derived stem cells differentiate to neuron-like cells in vitro

目的:研究脂肪来源干细胞(ASCs)的分离、纯化、扩增以及在体外定向分化为神经元样细胞的能力。方法:获取并培养正常人的ASCs,倒置显微镜下观察其形态;流式细胞仪检测其免疫表型。全反式维甲酸(ATRA)诱导ASCs在体外分化为神经元样细胞,倒置显微镜观察神经元样细胞的形态,逆转录-聚合酶链反应(RT-PCR)检测巢蛋白基因的表达;免疫组织化学染色法和Western blotting法检测神经丝蛋白(NF)和神经元烯醇化酶(NSE)的表达。结果:ASCs呈纤维样贴壁生长,体外培养易扩增;ASCs表达相关抗原CD29和CD105,不表达CD31、CD34、CD45和HLA-DR;不同扩增代数的ASCs经诱导剂的作用可呈现典型的神经元样细胞形态,巢蛋白基因及NF、NSE均呈阳性表达。结论:脂肪组织中分离培养的ASCs具有体外大量扩增并保持低分化状态的特性,以及定向分化为神经元样细胞的能力,是一种可用于神经系统疾病治疗的种子细胞。

AIM： We isolated and purified adipose - derived stem cells （ASCs） from adipose tissue in order to study their characters and examine their neuron differentiation capacity in vitro. METHODS. ASCs were isolated and cultured. The morphology of ASCs was observed under microscope and their phenotype was examined by flow cytometry. All - trans retinoic acid （ATRA） was used to induce ASCs into neuron -like cells. RT- PCR was used to detect the expression of nestin. Immunohistochemistry and Western blotting were used to detect the expression of neurofilament （NF） and neuron specific enolase （NSE）, respectively. RESULTS： ASCs displayed a fibroblast -like morphology adhering to the culture plate. The cells expressed several surface antigens such as CD29 and CD105, while did not express CD31, CD34, CD45 and HLA - DR. Under suitable conditions, various passages of ASCs all had the capacity of neuron differentiation. They expressed nestin, NF and NSE 10 days after adding ATRA in the culture system. CONCLUSION： ASCs isolated from adipose tissue can differentiate into neuron - like cells in vitro. ASCs may be used as an alternative source of cells for neural disease therapy.