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

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。

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

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 lymp-hocytes 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 co-cultured 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 bet-ween 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.