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Mitf和Pax6基因在小鼠胚胎干细胞向RPE细胞分化过程中的表达

, PP. 827-832

Keywords: 胚胎干细胞,分化,视网膜色素上皮细胞,Mitf,Pax

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Abstract:

目的探讨Mitf和Pax6基因在小鼠胚胎干细胞(mouseembryonicstemcells,mESCs)向视网膜色素上皮(retinalpigmentepithelium,RPE)细胞分化过程中的表达规律及其作用。方法免疫荧光染色、流式细胞仪检测分化前mESCs株在不同代数的干性标志表达情况。采用悬浮培养加诱导剂形成拟胚体(embryoidbodies,EBs)的方式诱导mESCs向RPE细胞分化,在分化前第0天,分化后第10、40天,采用流式细胞仪检测Mitf和Pax6在细胞中的表达规律及其与mESCs向RPE细胞分化的关系。结果mESCs株AB1在持续传代过程中形态一致且其干性标志SSEA1表达比例在不同代数之间差异无统计学意义(P>0.05)。分化所得细胞的RPE细胞成熟标志Bestropih、CRALBP、RPE65的表达比例分别为(61.6±2.4)%、(31.0±3.9)%、(57.8±2.0)%。在分化前第0天,Mitf、Pax6的表达比例分别为(1.7±1.5)%、(6.5±0.2)%,分化第10天分别为(28.5±7.2)%、(60.2±15.6)%,分化第40天分别为(5.7±2.1)%、(4.6±0.6)%。结论在mESCs向RPE细胞的分化过程中,Mitf和Pax6均呈先上升后下降的趋势,提示Mitf和Pax6参与了RPE细胞的分化过程。

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