体外传代培养对人视网膜色素上皮细胞免疫调控能力的影响  

Influence of serial subcultivation on the immunoregulation of retinal pigment epithelialium cells in vitro

作　　者：徐国兴[1] 李琼[1] 陈瑾[1] 郑学栋[1] 郭健[1] 谢茂松[1] 

机构地区：[1]福建医科大学附属第一医院眼科福建省眼科研究所,中国福建省福州市350005

出　　处：《国际眼科杂志》2008年第9期1777-1780,共4页International Eye Science

基　　金：中国卫生部科研基金资助项目(No.WKJ2005-2-013);中国福建省重大科研项目(No.002Y002);中国福建省福州市;福建医科大学教授发展基金课题(No.2006-js6033)~~

摘　　要：目的:研究体外传代培养对人类视网膜色素上皮(human reti-nal pigment epithelialium,HRPE)细胞免疫调控能力的影响。方法:建立HRPE细胞与异体淋巴细胞共培养系统,采用流式细胞术检测原代及传代HRPE细胞(第1、2、3、4、5、6代)Fas表达以及诱导异体淋巴细胞凋亡能力的差异。结果:HRPE与淋巴细胞体外共培养时,HRPE细胞Fas表达随着传代逐渐减弱,Fas表达与传代代数呈负相关(阳性细胞率r=-0.859,P<0.01;平均荧光强度r=-0.880,P<0.01),HRPE细胞诱导淋巴细胞凋亡随着传代逐渐减弱,淋巴细胞凋亡率与传代代数呈负相关(r=-0.838,P<0.01),共培养时传代HRPE细胞Fas表达迅速下降,至第3代与第4代间Fas表达差异无统计学意义(P>0.05),共培养时传代HRPE细胞诱导淋巴细胞凋亡率迅速下降,至第3代与第4代间淋巴细胞凋亡率差异无统计学意义(P>0.05)。结论:体外传代培养的HRPE细胞,传代数越大细胞变异越大,第3代后细胞的免疫调控能力明显下降。AIM ： To investigate the influence of serial subcultivation on the immunoregulation of retinal pigment epithelialium cells in vitro . METHODS： Co-culture-systems of HRPE cells with lymphocytes were established in vitro . The expression of Fas on different passages of HRPE cells and the induction of apoptosis on lymphocytes were observed by flow cytometry. RESULTS： When coultured with lymphocytes, the expression of Fas was tapering and the duration of culture in vitro were significantly correlated with the expression of Fas （the rate of positive cell： r = -0.859, P 〈 0.01, the mean fluorescence intensity： r=-0.880,P〈0.01 ）. The rate of apoptosis of lymphocytes was taper going down to posterity, and the duration of culture in vitro were significantly correlated with the rate of apoptosis of lymphocytes（ r= -0.838, P〈 0.01 ）. The expression of Fas and the rate of apoptosis of lymphocytes quickly descend going down to posterity. There was no difference between the third passage and the fourth passage （P〉0.05）. CONCLUSION： HRPE cells beyond 3 passages show a low capability of immunoregulation, and are not suitable for seeds cells for HRPE transplantation.

关 键 词：视网膜色素上皮细胞 传代培养 淋巴细胞 凋亡 

分 类 号：R774.1[医药卫生—眼科] R331.22[医药卫生—临床医学]