中文摘要：

目的：探讨细胞生长因子联合诱导胚胎干细胞（embryonicstemcell，Es细胞）向肾脏前体细胞定向分化的实验技术，为应用胚胎干细胞进行肾脏再生提供实验基础。方法：小鼠ES细胞悬浮培养2d制备成拟胚体，RA、activin—A、Bmp7三种生长因子诱导培养5d设为A组，不加生长因子培养设为对照（B组），生长因子培养5天后继续以肾脏上皮细胞生长培养液培养7d设为c组，免疫荧光检测Pax2、Bry、wTl蛋白表达情况，流式细胞技术检测Pax2、Bry、cD24阳性细胞比例，realtimePcR检测Pa]（2、Bry、0srl、Liml、wTl、six2、salll、AQPl、CD24、PDGFR基因表达情况。结果：免疫荧光染色结果显示A组细胞Pax2、Bry、WTl的表达明显高于B组；流式细胞技术检测显示A组Pax2、Bry、cD24阳性细胞比例较B组增加，C组表达Pax2、Bry的阳性细胞比例较A组明显升高；realtimePcR检测显示A组Pax2、Bry、0srl、Ijml、wTl表达较B组明显增加（P〈0．05或P〈0．01），C组表达CD。基因较A组明显升高（P〈0．05）。结论：胚胎干细胞在体外生长因子联合诱导条件下能够分化为早期肾脏前体细胞，并表达部分成熟肾脏细胞的标记物。

英文摘要：

Objective：To investigate whether ES cells can be induced into renal progenitor ceils in vitro with cell growth factors, and provide the experimental basis for the application of embryonic stem cells in kidney regeneration. Methods： The dissociated mouse ES ceils formed embryoid bodies in suspension culture. EB was treated with a combination of growth factors （ RA, activin - A, Bmp7）, and renal epithelial cell growth medium. After 5 days of induction, Pax2, Bry, WT1 were detected in EB cultured by immu- nofluorescence staining. Flow Cytometry analyzed the proportion of Pax2 ＋ , Bry ＋ and CD24 ＋ cells. Real time PCR analyzed the renal developmentally regulated genes Pax2, Bry, Osrl, Liml, WT1, Six2, Salll, AQP1, CD24, PDGFR. Results： Iimmunofluorescence stai- ning showed that the expression of Pax2, Bry, WT1 were higher in A group compared with B group. Flow Cytometry detected that the proportion of Pax2 ＋, Bry ＋ and CD24 ＋ cells in A group was higher than that in B group, and the proportion of Pax2 ＋, Bry ＋ cells in C group was significantly higher than that in A group. Real time PCR analysis showed that the Pax2 ,Bry,Osrl ,Liml ,WT1 genes were significantly increased in A group compared with B group（ P 〈 0.05 or P 〈0.01 ）, and showed that the CD24 genes were signifi- cantly increased in C group compared with A group （ P 〈 0.05 ）. Conclusion： Mouse ES cells can be induced into renal progenitor cells with a combination of growth factors, and express partial mature kidney cells markers.