多因子联合诱导脂肪组织源性干细胞分化为神经元样细胞

背景：国内均可见有探讨脂肪干细胞向神经元诱导分化的相关研究报道，但运用差速贴壁法纯化脂肪组织源性干细胞，多因子联合诱导脂肪组织源性干细胞分化为神经元样细胞的研究鲜有报道。 目的：观察多因子联合诱导和分步诱导方案诱导大鼠脂肪组织源性干细胞体外分化为神经元样细胞的可行性。 方法：取SD雄性大鼠腹股沟和附睾脂肪垫脂肪组织，酶消化法分离、差速贴壁法纯化脂肪组织源性干细胞。免疫荧光细胞化学法鉴定脂肪组织源性干细胞(CD44，CD49d，CD106)；“鸡尾酒”式神经元诱导培养基对脂肪组织源性干细胞进行诱导分化，并用免疫荧光细胞化学法鉴定(Nestin和NeuN)分化结果。 结果与结论：大鼠腹股沟和附睾脂肪垫脂肪组织内分离、原代培养的脂肪组织源性干细胞形态均一，呈长梭形，细胞核椭圆形，核仁明显，细胞质染色浅，呈旋涡状克隆样生长；CD44和CD49d阳性表达而CD106阴性；诱导培养基内培养6 h后脂肪组织源性干细胞呈锥体形或不规则，有短而细的指状突起；24 h后多数细胞显示类神经元的形态学特征，且Nestin和NeuN阳性。提示肪组织源性干细胞在多因子“鸡尾酒”式培养基内可向神经元方向分化。

Multiple factors-induced differentiation of rat adipose tissue-derived stem cells into neuron-like cells

BACKGROUND: Differentiation of adipose tissue-derived stem cells (ADSCs) into neurons has been reported. However, little data are available regarding differential adhesion-purified ADSCs differentiation into neuron-like cells induced by multiple factors. OBJECTIVE: To explore the feasibility of inducing rat ADSCs into neuron-like cells with a new scheme and medium containing multiple factors. METHODS: ADSCs were isolated from the fat pads in groins and epididymis of male Sprague Dawley rats by enzyme digestion, and purified by differential attachment method. Fluorescent immunocytochemistry was used to identify the phenotype of the cells (CD44, CD49d and CD106 expressions). Differentiation of ADSCs into neuron-like cells was induced in cocktail-like neuronal differential medium containing insulin, hydrocortisone, valproic acid and forskolin. Also, fluorescent immunocytochemistry was used to identify the markers of the differentiated cells (Nestin and NeuN). RESULTS AND CONCLUSION: After sequentially isolated, purified and cultured, the obtained cells presented with long-fusiform shape, with clear nucleoli and light stained cytoplasm. They were positive for CD44 and CD49d, but negative for CD106. The application of NIM resulted in morphological changes of ADSCs during the first 6 hours, shown by pyramidal, irregular or triangular cell body with short, thin finger-like cell processes. After 24 hours of induction, most of the cells developed multiple branched processes resembling those of neurons and positive for Nestin and NeuN. Results showed that ADSCs could be induced to differentiate into neuron-like cells in cocktail-like neuronal differential medium.