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] 

机构地区：[1]福建医科大学附属第一医院/福建省眼科研究所,福州350005

出　　处：《医学研究杂志》2009年第1期26-29,F0003,共5页Journal of Medical Research

基　　金：国家人事部留学回国人员优秀课题基金资助(Rsbkj2006-1);福建省重大科学研究课题基金资助(2002Y002)

摘　　要：目的研究体外传代培养对人类视网膜色素上皮(human retinal 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<0.001;平均荧光强度r=-0.880,P<0.001),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<0.001),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。Objective 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 differ- ent passages of HRPE cells and the induction of apoptosis on lymphocytes were observed by flow cytometry. Results When co - cultured with lymphocytes,the expression of Fas was tapering and the duration of culture in vitro was significantly correlated with the expression of Fas（ the rate of positive cell： r = -0. 859,P 〈 O. 001 ; the mean fluorescence intensity： r = -0. 880 ,P 〈 0. 001 ）. The rate of apoptosis of lymphocytes was taper going down to posterity,and the culture in vitro was significantly correlated with the rate of apoptosis of lymphocytes （r = -0. 838 ,P 〈 0. 001 ）. 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 showed a low capability of immunoregulation and were not suitable for seeds ceils 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]