论文部分内容阅读
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6h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;24h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。
BACKGROUND: There are many studies on the differentiation of adipose derived stem cells into neurons in China. However, adipose tissue-derived stem cells are differentiated into neuron-like cells by differential adherent method. Little report. OBJECTIVE: To observe the feasibility of induction of adipose derived stem cells into neuron-like cells in vitro by multi-factor combined induction and stepwise induction. METHODS: SD rats were injected with adipose tissue of the inguinal and epididymal fat pad, isolated by enzymatic digestion and purified by adipose tissue-derived stem cells. Adipose tissue-derived stem cells (CD44, CD49d, CD106) were identified by immunofluorescence cytochemistry; adipose tissue-derived stem cells were induced to differentiate by “cocktail” neuronal induction medium and identified by immunofluorescence cytochemistry (Nestin and NeuN) differentiation results. RESULTS AND CONCLUSION: The adipose tissue-derived stem cells isolated from the inguinal and epididymal fat pad were homogenized in the form of long spindle, oval nucleus, obvious nucleoli and shallow cytoplasm, Growth; CD44 and CD49d positive expression and CD106 negative; induced adipose tissue-derived stem cells were pyramidal or irregular after induction for 6h, with short and thin finger-like protrusions; most of the cells showed morphology of neurons after 24h Features, and Nestin and NeuN positive. It indicated that adipose tissue-derived stem cells can differentiate into neurons in multi-factor “cocktail” medium.