羊膜上皮细胞条件培养液对大鼠骨髓基质细胞神经分化的影响

THE EFFECTS OF AECs CONDITIONED MEDIUM ON INDUCING NEURONAL DIFFERENTIATION OF BMSCs

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 study the effects of amniotic epithelial cells conditioned medium on the differentiation of bone marrow stromal cells into neural cells. Methods Bone marrow stromal cells and amniotic epithelial cells were isolated and cultured in vitro, then the cell surface antigen was detected by flow cytometry and the expressions of nestin and ki67 were detected by immunofluorescence staining method. When the cells were co-cultured with amniotic epithelial cells conditioned medium, the morphological character of ceils was observed by inverse phase-contrast microscope, and the expressions of NSE（ neurone specific enolase）, TH （tyrosine hydroxylase ） and DAT（ dopamine transporter） were detected by immunofluorescence staining method. Results Amniotic epithelial cells conditioned medium had obvious inductive effect on bone marrow stremal cell＇s neural differentiation. Conclusion The amniotic epithelial ceils conditioned medium may have inductive effect neuron-like cell＇s differentiation and dopaminergic neuron-like cell＇s differentiation of bone marrow stromal cells in vitro.