不同浓度维甲酸诱导体外培养人羊膜上皮细胞分化为神经样细胞的研究

目的探讨维甲酸诱导人羊膜上皮细胞向神经样细胞分化。方法以DMEM/F12培养基培养原代人羊膜上皮细胞,并将细胞分为5组:对照组;1×10-8mol·L-1维甲酸组;1×10-7mol·L-1维甲酸组;1×10-6mol·L-1维甲酸组;1×10-5mol·L-1维甲酸组。各维甲酸组经维甲酸处理7 d后,用免疫组化检测神经干细胞巢蛋白(nestin)、微管相关蛋白2(MAP-2)、神经胶质纤维酸性蛋白(GFAP)和细胞全能性标记物Oct-4表达,并计数阳性细胞比例。结果与对照组比较,各维甲酸组显著提高人羊膜上皮细胞分化为神经样细胞的比例,其中1×10-6mol·L-1维甲酸组阳性细胞比例最高;结论维甲酸体外能诱导人羊膜上皮细胞向神经样细胞分化,最适诱导浓度为1×10-6mol·L-1。

Research on Inducing Human Amnionic Epithelial Cells Differentiate into Neural-like Cells with Different Concentration of Retinoic Acid in Vitro

Aim To investigate the effect of retinoic acid （RA） on the human amnionic epithelial cells.Methods The human amnionic epithelial cells are cultured in DMEM/F12 medium in vitro.The cells were divided into five groups：① Control group; ②（1 × 10-8） mol·L-1 RA group：the cells treated with DMEM/F12 medium containing （1 × 10-8）mol·L-1 RA; ③（1 × 10-7）mol·L-1 RA group：the cells treated with DMEM/F12 medium containing （1 × 10-7）mol·L-1 RA; ④（1 × 10-6）mol·L-1 RA group：the cells treated with DMEM/F12 medium containing （1 × 10-6）mol·L-1 RA; ⑤（1 × 10-5）mol·L-1 RA group：the cells treated with DMEM/F12 medium containing （1 × 10-5）mol·L-1 RA.After 7 days,the expression of Oct-4,Nestin,MAP-2 and GFAP in the human amnionic epithelial cells were investigated by immunohistochemical technology.Results As immunohistochemical technology and Western-blotting technology showed,the expression ofnestin,MAP-2 and GFAP were higher in the other groups than in the control group,which were the most in （1 × 10-6）mol·L-1 RA group.The expression of Oct-4 was less in the other groups than the control group,which were the least in （1 × 10-6）mol·L-1 RA group.Conclusion RA can increase the expression of Nestin,MAP-2 and GFAP,while decrease the expression of Oct-4,which means that RA can induce the human amnionic epithelial cells differentiate into neural-like cells in vitro.And （1 × 10-6）mol·L-1 RA is the appropriate concentration.