Department of Ophthalmology, Peking University Third Hospital, Key Laboratory of Vision Loss and Restoration, Ministry of Education, Beijing, People's Republic of China.
Am J Ophthalmol. 2013 Mar;155(3):474-483.e2. doi: 10.1016/j.ajo.2012.09.010. Epub 2012 Dec 4.
To evaluate the morphologic features and viability of human retinal pigment epithelium (RPE) cells captured from the pigment epithelium detachment (PED) region outside the choroidal neovascular membrane lesion in eyes with hemorrhagic age-related macular degeneration.
Prospective, observational case series.
Five specimens of the RPE sheet were obtained from the PED region after choroidal neovascular membrane excision in 5 eyes of 5 patients during RPE transplantation for hemorrhagic age-related macular degeneration. The specimens were stained with hematoxylin and eosin and with periodic acid-Schiff. Immunohistochemistry analysis for RPE-65 and zonula occludens-1 also was performed. RPE cells from the PED region were cultured and passaged 5 times. Scanning and transmission electron microscopy were performed to analyze the specimens.
The RPE cells of the specimens contained brownish pigment and were autofluorescent in vitro. Periodic acid-Schiff staining revealed that the Bruch membrane below the RPE monolayer was not intact. The specimens demonstrated positive results for both zonula occludens-1 and RPE-65 staining. The RPE basement membrane in the specimen was observed by both scanning and transmission electron microscopy. Intercellular tight junctions among RPE cells of the specimen also were observed. RPE cells captured from the PED region were cultured successfully and were passaged 5 times.
The RPE sheet developed from the PED region outside the choroidal neovascular membrane lesion had tight intercellular junctions, a simple RPE basement membrane, and active cellular viability. This monolayer RPE sheet may be considered as a substitution for subfoveal RPE loss in eyes with hemorrhagic age-related macular degeneration.
评估从伴有出血性年龄相关性黄斑变性脉络膜新生血管膜病变之外的色素上皮脱离（PED）区域捕获的人视网膜色素上皮（RPE）细胞的形态特征和活力。
前瞻性、观察性病例系列。
在 5 例患者的 5 只眼中，在进行脉络膜新生血管膜切除后，从PED 区域获得了 5 个 RPE 片标本，用于因出血性年龄相关性黄斑变性进行的 RPE 移植。用苏木精和伊红以及过碘酸-Schiff 染色对标本进行染色。还进行了 RPE-65 和封闭蛋白-1 的免疫组织化学分析。从 PED 区域分离的 RPE 细胞培养和传代 5 次。进行扫描和透射电镜检查以分析标本。
标本中的 RPE 细胞含有棕褐色色素，在体外具有自发荧光。过碘酸-Schiff 染色显示，RPE 单层下方的 Bruch 膜不完整。标本对封闭蛋白-1 和 RPE-65 染色均呈阳性结果。扫描和透射电镜观察到标本中的 RPE 基膜。还观察到标本中 RPE 细胞之间的细胞间紧密连接。从 PED 区域捕获的 RPE 细胞成功培养并传代 5 次。
从脉络膜新生血管膜病变之外的 PED 区域发育而来的 RPE 片具有紧密的细胞间连接、简单的 RPE 基膜和活跃的细胞活力。这种单层 RPE 片可被视为替代伴有出血性年龄相关性黄斑变性的黄斑下 RPE 丧失的替代物。
