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目的:观察大鼠嗅鞘细胞无血清上清液诱导脐血间充质干细胞向神经细胞分化的作用。方法:采用差速贴壁联合阿糖胞苷抑制法获得较高纯度的嗅鞘细胞,在最佳状态细胞无血清培养后取上清液。收集正常足月剖腹产胎儿的脐带血,经肝素抗凝,用淋巴细胞分离液分离脐血的单个核细胞,采用差速贴壁法纯化间充质干细胞,传第三代后用上述收集的嗅鞘细胞无血清上清液诱导,观察诱导分化过程,免疫组化鉴定分化结果。结果:用差速贴壁和阿糖胞苷抑制法可获得纯度高达95%的嗅鞘细胞,使用无血清嗅鞘细胞上清液培养脐血间充质干细胞后,发现能诱导脐血间充质干细胞向神经细胞形态分化,胞体呈椭圆形,伸出长突起。诱导7天后相差显微镜下可观察到神经干细胞、神经元和胶质细胞形态的细胞,与NSE、GFAP细胞免疫组化结果一致。结论:大鼠嗅鞘细胞无血清上清液有明显诱导脐血间充质干细胞分化为神经细胞的作用。
OBJECTIVE: To observe the effect of serum-free supernatant of rat OECs on differentiation of umbilical cord blood mesenchymal stem cells into neurons. Methods: Olfactory ensheathing cells were obtained by differential adhesion and cytarabine inhibition. Supernatants were obtained after serum-free culture in the optimal condition. Umbilical cord blood was collected from normal full-term cesarean section, isolated from umbilical cord blood by lymphocyte separation by heparin anticoagulation, purified mesenchymal stem cells by differential adherent method, Sheath cells were induced by serum-free supernatant, the differentiation process was observed, and the differentiation results were identified by immunohistochemistry. RESULTS: Olfactory ensheathing cells with a purity of up to 95% were obtained by differential adherence and cytarabine inhibition. After cord blood-derived mesenchymal stem cells were cultured with serum-free olfactory ensheathing cell supernatant, they were found to induce cord blood filling Mature stem cells differentiate into neuronal cells, and the cell bodies are oval in shape and extend into long protrusions. After 7 days of induction, the morphological changes of NSCs, neurons and glial cells were observed under the phase contrast microscope, which was consistent with the results of immunohistochemistry of NSE and GFAP cells. Conclusion: The serum-free supernatant from rat OECs can induce the differentiation of UCB-MSCs into neurons.