活体染料标记原代IPEC眼内移植的研究  

Intraocular transplantation of iris pigment epithelial cells labelled with vital dye

作　　者：孙大卫[1] 鲍延丽[1] 焦万秋[1] 彭绍民[1] 

机构地区：[1]哈尔滨医科大学第二临床医学院眼科,黑龙江哈尔滨150081

出　　处：《哈尔滨医科大学学报》2007年第3期202-205,共4页Journal of Harbin Medical University

基　　金：国家自然科学基金资助项目(30271395);哈尔滨医科大学第二临床医学院青年基金(QN200603)

摘　　要：目的探讨应用活体染料羧基荧光素乙酰乙酸(CFDA-SE)标记原代的兔自体虹膜色素上皮细胞(iris pig-ment epithelial cell,IPEC),并移植于视网膜神经上皮层下的可行性。方法取原代兔眼自体虹膜色素上皮细胞消化分离,经CFDA-SE染色后移植到视网膜神经上皮层下。术后不同时间应用彩色眼底照相、激光扫描检眼镜(SLO)、光学相干断层扫描(OCT)进行活体观察,不同时间点对移植眼进行切片并常规染色及免疫组化,用光学显微镜对移植细胞进行观察和评价。结果自体移植术后观察显示移植区界限清晰;SLO可观察到移植区内荧光分布;OCT和HE染色均显示移植区神经上皮层下有移植物堆积,并随时间推移逐渐变平。免疫组化显示移植区ZO-1、Actin呈棕色点,有规律地间断分布在Bruch膜上,IPEC嵌插在原始的视网膜色素上皮细胞(retinal pigment epithelial cell,RPEC)间,且形成完整的单层。结论CFDA-SE作为活体染料标记自体IPEC进行移植时,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。Objective To investigate the feasibility of application of vital dye carboxyfluorescein diacetate Succinimidyl Ester （CFDA-SE） to label rabbit primary iris pigment epithelial cells （IPEC） for transplantation into subretinal space.Methods Isolated rabbit IPECs and labelled them with CFDA-SE, then transplanted them into subretinal space. The transplanted IPECs were observed by color fundus photography, fundus fluorescein angiography （FFA）, and optical coherence tomography （OCT）. In addition, cellular configuration and tight junction protein were examined by light and fluorescence microscopy and immunohistochemistry. Results The rabbit primary IPEC labeled by CFDA-SE could be observed clearly in the subretinal space. Through SLO and OCT, it was observed that the primary IPEC formed one or more contiguous layers on top of native RPE in the earlier days.Mter 3 months,grafted IPEC were present in the subrefinal space, often forming monolayer-like chains integrating with the native RPEC. Immunostaining experiments showed that IPECs survived and compatible with RPEC, and formed tight junctions. Conclusion CFDA-SE is a kind of stable and safe vital dye to label IPEC for tracing of transplantation.

关 键 词：活体染料 虹膜色素上皮细胞 细胞移植 原代细胞 

分 类 号：R774[医药卫生—眼科]