摘要
目的探讨兔眼自体虹膜色素上皮细胞(iris pigment epithelial cells,IPECs)羧基荧光素乙酰乙酸(CFDA-SE)标记后,采用单通道内路移植方法将传代的IPECs移植于视网膜神经上皮层下的可行性。方法取兔眼自体虹膜细胞分离培养并传代,于第3代经CFDA-SE染色后移植到自体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、光学相干断层扫描进行活体观察,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。实验数据采用两均数的等效检验,以0.5倍标准差作为等效界值观察移植前后ZO-1和Actin表达情况。结果移植术后观察显示移植区界限清晰;激光扫描检眼镜可观察到移植区内荧光;光学相干断层扫描和HE染色均显示移植区神经上皮层下有色素上皮细胞堆积,并随时间推移堆积逐渐变平。经免疫组化显示移植区ZO-1、Actin呈棕色点,有规律的间断分布在Bruch膜上,IPECs嵌插在原始的RPECs间,且形成完整的单层。结论CFDA-SE是一种快捷、稳定、安全的活体细胞染料,在IPECs移植中可作为理想的标记物应用;我们首次使用的单通道内路移植方法具有手术快捷简单、安全、并发症少等优点。
Objective To investigate the feasibility of application of vital dye CFDA-SE to label autologous iris pigment epithelial cells （IPECs） in auto- plastic transplantation into subretinal space. Methods Autologous IPECs were cultured and passagcd to the third generation,labeled with CFDA-SE and then transplanted into subretinal space. Viviperception was performed by color fundus photography, scanning laser ophthalmoscope and optical coherence tomography （OCT） at different time after the operation. In addition,cellular configuration and tight junction protein were examined with light and fluorescence microseopy and immtmohistochemistry. The experimental data was analyzed by iso-effective test, 0. 5 double standard deviation was set as the iso-effective group value to observe the expression of ZO-1 and Actin before and after the txansplantation. Results The autologous IPECs labeled by CFDA-SE could be observed clearly in the subretinal space, where the fluorescence Could be observed by scanning laser ophthalmoscope and the cumulated pigment epithelial cells which flattened gradtually with the time prolonged were showed by optical coherence tomography and HE staining. The results of immtmohistochemtstry showed that ZO-1 and Actin in the subretinal space were presented as brown points and distributed on the Bruch membrane regularly and discontinuously. The IPECs inserted in the native retinal pigment epithelial cells and formed single layer. Immunostaining experiments showed that IPECS survived and compacted with retinal pigment epithelial cells, and formed tight junctions.Conclusion CFDA-SE is a land of stable and safety vital dye to label IPECs in txansplantation. The method we used to txansplant cells into subretianl space is effective and accompanied less complications. [Rec Adv Oohtbalmol 2007;27（3） ：165-169]
出处
《眼科新进展》
CAS
2007年第3期165-169,共5页
Recent Advances in Ophthalmology
基金
国家自然科学基金资助(编号:30271395)
黑龙江省海外学人科研基金资助(编号:1053HQ014)
哈尔滨医科大学附属第二医院青年基金资助(编号:QN200603)~~
关键词
虹膜
色素上皮
上皮细胞
活体染料
细胞移植
兔
iris
pigment epithelial
epithelial cells
vital dye
cell transplantation
rabbit
