视网膜色素上皮细胞移植中CFDA-SE及免疫组化研究  

Study on CFDA-SE and immunohistochemistry in the transplantation of retinal pigment epithelium cells

作　　者：刘春艳[1] 孙大卫[1] 宁静[1] 彭绍民[1] 

机构地区：[1]哈尔滨医科大学附属第二临床医学院眼科,150086

出　　处：《眼科新进展》2006年第5期321-323,共3页Recent Advances in Ophthalmology

基　　金：国家自然科学基金资助(编号:30271395);黑龙江省教育厅海外学人科研基金资助;哈尔滨医科大学研究生创新基金资助

摘　　要：目的探讨新型染料羧基荧光素乙酰乙酸[5-(and-6)-carboxyfluresceindiacetate,succinimidylester,CFDA-SE]与免疫组化方法相结合在视网膜色素上皮细胞(retinalpig-mentepithelialcells,RPECs)移植中应用的可行性。方法用CFDA-SE(10μmol·L-1)标记供体RPECs,37℃下孵育1min,通过玻璃体视网膜显微手术将细胞悬液植入同种异体青紫蓝兔视网膜下腔,分别在30d和60d时处死实验兔,荧光显微镜下观察标记的供体细胞,同时作紧密连接的结构蛋白ZO-1和细胞骨架蛋白Actin的免疫组化分析。结果荧光显微镜下观察到标记的供体RPECs可嵌插于宿主RPECs之间,铺成单层结构,并形成新的细胞间紧密连接,ZO-1、Actin的免疫组化结果经统计分析表明与原位RPECs之间的表达无显著差异(ZO-1:︱t︱=2.05,P<0.05;Actin:︱t︱=2·14,P<0.05)。结论CFDA-SE是一种快捷、稳定、安全的活体细胞染料,可作为理想的标记物应用于RPECs移植中。Objective To test and verify the feasibility of application of vital dye CFDA-SE to label retinal pigment epithelial cells（RPECs） in transplantation of retinal pigment epithelium. Methodhs Allogenic pigmented rabbit RPECs were labeled with 10 pmol· L^-1 CFDA-SE for 1 minute at 37 ℃, then washed and transplanted into the subretinal space of the fellow eye balls using vitreous body retina microsurgery. Rabbits were killed 30 days or 60 days after the implantation,and immunostaining for the expression of structure protein ZO-1 and cell skeleton protein Actin were performed. The light and fluoresent microscopy were applicated to examine the survival of the labeled cells. Resuits Immunostaining experiments showed that labeled RPECs survived and compacted among the host RPECs, formed tight junctions and a polarized monolayer with the native RPECs. Statistical analysis showed it has no significant difference which RPECs in situ（ZO-1： ｜ t ｜ =2.05,P（0.05;Actin： ｜ t ｜ = 2.14,P（0.05）.Conclusion The fluorescent dye CFDA-SE can be used as a new marker of rabbit RPECs in transplantation due to its characteristic of safty, convenience and nontoxity.

关 键 词：视网膜色素上皮细胞 羧基荧光素乙酰乙酸 移植 免疫组化 

分 类 号：R774.1[医药卫生—眼科]