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目的探讨羊膜上皮细胞条件培养液对大鼠骨髓基质细胞分化为神经细胞的作用。方法体外分离、培养羊膜上皮细胞及大鼠骨髓基质干细胞,流式细胞术检测其表面抗原,免疫组织化学染色技术检测巢蛋白(Nestin)以及增殖细胞相关核抗原(ki67)在细胞中的表达。采用羊膜上皮细胞条件培养液对其进行诱导,倒置相差显微镜观察细胞形态的变化,免疫荧光染色技术检测神经特异烯醇化酶(NSE)以及多巴胺能神经元标志物酪氨酸羟化酶(TH)和多巴胺转运体(DAT)在诱导后细胞中的表达,并进行定量比较分析。结果羊膜上皮细胞条件培养液能够诱导大鼠骨髓基质细胞向神经细胞方向分化。结论羊膜上皮细胞条件培养液可以诱导大鼠骨髓基质细胞分化为神经元样细胞和多巴胺能神经元样细胞。
Objective To investigate the effect of amniotic epithelial conditioned medium on the differentiation of rat bone marrow stromal cells into neurons. Methods Human amniotic epithelial cells and rat bone marrow stromal cells were isolated and cultured in vitro. The surface antigens were detected by flow cytometry. The expressions of Nestin and proliferating cell nuclear antigen (ki67) were detected by immunohistochemical staining. The cells were cultured in conditioned medium containing amniotic epithelial cells, the morphological changes of cells were observed by inverted phase contrast microscope, and the activity of neuron specific enolase (NSE) and tyrosine hydroxylase (TH), a marker of dopaminergic neuron, were detected by immunofluorescence staining. And dopamine transporter (DAT) in the cells after induction of expression and quantitative comparative analysis. Results Conditioned medium of amniotic epithelial cells induced the differentiation of rat bone marrow stromal cells to nerve cells. Conclusion Amniotic epithelial cell conditioned medium can induce rat bone marrow stromal cells to differentiate into neuron-like cells and dopaminergic neuron-like cells.