發(fā)布時(shí)間：2018-12-12 14:25

【摘要】：目的:通過(guò)誘導(dǎo)多能干細(xì)胞(iPSCs)技術(shù)獲得縫隙連接蛋白26(Cx26)缺陷聾患兒的神經(jīng)細(xì)胞,研究Cx26缺陷對(duì)神經(jīng)細(xì)胞發(fā)育和基因表達(dá)的影響。方法:從3例Cx26缺陷造成的極重度耳聾患兒取得成纖維細(xì)胞,將之誘導(dǎo)為數(shù)個(gè)非整合iPSC細(xì)胞系,鑒定這些細(xì)胞系的形態(tài)和內(nèi)源性、外源性基因表達(dá)。然后將這些細(xì)胞系向神經(jīng)細(xì)胞方向分化,檢測(cè)整個(gè)過(guò)程中形態(tài)、多能性基因、神經(jīng)標(biāo)記物、縫隙連接蛋白基因表達(dá)變化。結(jié)果:能夠成功建立Cx26缺陷的3個(gè)iPSC細(xì)胞系,并且可以在體外分化為神經(jīng)前體細(xì)胞和神經(jīng)元細(xì)胞;這些細(xì)胞的形態(tài)、增殖、內(nèi)源性、外源性基因表達(dá)與人胚胎干細(xì)胞基本一致;iPSC細(xì)胞系分化出的神經(jīng)元細(xì)胞中,Cx32的表達(dá)明顯上調(diào),Cx36的表達(dá)略微上調(diào),Cx26的表達(dá)沒(méi)有明顯變化。結(jié)論:Cx26缺陷不影響誘導(dǎo)多能干細(xì)胞的神經(jīng)分化,但其過(guò)程中Cx32和Cx36表達(dá)上調(diào),提示Cx32可能對(duì)Cx26缺陷發(fā)揮了代償作用。
[Abstract]:Aim: to obtain nerve cells from deafness children with gap junction protein 26 (Cx26) deficiency by inducing pluripotent stem cell (iPSCs), and to study the effect of Cx26 deficiency on neuronal development and gene expression. Methods: fibroblasts were obtained from 3 children with extremely severe deafness caused by Cx26 deficiency. The fibroblasts were induced into several unintegrated iPSC cell lines and their morphology, endogenous and exogenous gene expression were identified. Then these cell lines were differentiated into nerve cells, and the expression changes of morphology, pluripotent genes, neural markers and gap junction protein genes in the whole process were detected. Results: three iPSC cell lines with Cx26 deficiency could be successfully established and differentiated into neural precursor cells and neuron cells in vitro, and the morphology, proliferation, endogenous and exogenous gene expression of these cells were basically consistent with those of human embryonic stem cells. In the differentiated neurons of iPSC cell line, the expression of Cx32 was up-regulated, the expression of Cx36 was slightly up-regulated, and the expression of Cx26 was not changed. Conclusion: Cx26 deficiency does not affect the neural differentiation of pluripotent stem cells, but the expression of Cx32 and Cx36 is up-regulated, suggesting that Cx32 may play a compensatory role on Cx26 deficiency.
【作者單位】： 中國(guó)醫(yī)學(xué)科學(xué)院北京協(xié)和醫(yī)院耳鼻咽喉科;中國(guó)醫(yī)學(xué)科學(xué)院基礎(chǔ)醫(yī)學(xué)研究所;
【分類(lèi)號(hào)】：R764.43

【相似文獻(xiàn)】

相關(guān)期刊論文 前1條

1 林英;鄭建秋;葛紅巖;侯麗麗;呂鋒;金迪;馬洪梅;付小玻;劉平;;縫隙連接蛋白50真核表達(dá)載體的構(gòu)建與表達(dá)[J];國(guó)際眼科雜志;2007年05期

相關(guān)會(huì)議論文 前2條

1 李慶忠;;縫隙連接蛋白分子流行病學(xué)研究[A];華東六省一市耳鼻咽喉-頭頸外科學(xué)術(shù)會(huì)議暨2008年浙江省耳鼻咽喉-頭頸外科學(xué)術(shù)年會(huì)論文匯編[C];2008年

2 郭書(shū)忍;鄭芳;劉媛媛;陳詠梅;;湖北省漢族人群中縫隙連接蛋白GJA8基因外顯子區(qū)-201T/C單核苷酸多態(tài)性分布特征研究[A];中華醫(yī)學(xué)會(huì)第七次全國(guó)檢驗(yàn)醫(yī)學(xué)學(xué)術(shù)會(huì)議資料匯編[C];2008年

，

本文編號(hào)：2374737