Impaired RPE survival on aged submacular human Bruch's membrane
Impaired RPE survival on aged submacular human Bruch's membrane
复制标题
DOI：
10.1016/j.exer.2004.09.006
复制
发表时间：
2005-02-01
影响因子：
3.4
通讯作者：
Zarbin, MA
中科院分区：
文献类型：
--
作者：
Gullapalli, VK;Sugino, IK;Zarbin, MA
已提出用健康的视网膜色素上皮（RPE）对患病或医源性损伤的布鲁赫膜进行表面置换，作为年龄相关性黄斑变性（AMD）的辅助治疗。本研究的目的是确定培养的人胚胎RPE细胞是否可以在老年人黄斑下Bruch膜上附着和分化。清除Bruch膜以暴露天然RPE基底膜、RPE基底膜正下方的浅表内胶原层或深部内胶原层。这是移植的RPE细胞将在原位遇到的三个表面。将大约3146个培养的胎儿RPE细胞mm（-2）接种到这三个表面上，并在器官培养物中生长1、7或14天。将外植体一分为二并进行组织学检查或用扫描电子显微镜分析。在染色切片上测量RPE核密度。将形态学和细胞密度与接种到牛角膜内皮细胞-细胞外基质（BCE-ECM）上的细胞进行比较。还在妊娠18周到88岁的供体眼睛的组织切片中测量了原位黄斑下RPE核密度，以确定年龄对RPE密度的影响。与以相似密度接种到BCE-ECM上的细胞相比，在所有时间点，老年人黄斑下布鲁赫膜上的RPE细胞覆盖率和细胞形态都很差。与BCE-ECM上的细胞相反，Bruch膜上的RPE细胞密度随时间而降低。一般来说，与第L天相比，所有三种Bruch膜表面上的细胞形态在第7天恶化。尽管在第7天一些细胞在RPE基底膜和深层内胶原层上色素沉着更多，但差的细胞形态表明剩余细胞没有很好地分化。在第14天，细胞在BCE-ECM上是均匀的和立方形的，细胞密度与第7天相似，与年轻供体的原位密度相似。
Resurfacing of diseased or iatrogenically damaged Bruch's membrane with healthy retinal pigment epithelium (RPE) has been proposed as adjunctive treatment for age-related macular degeneration (AMD). The purpose of this study was to determine whether cultured fetal human RPE cells can attach and differentiate on aged submacular human Bruch's membrane. Bruch's membrane was debrided to expose native RPE basement membrane, the superficial inner collagenous layer directly below the RPE basement membrane, or the deep inner collagenous layer. These are three surfaces that transplanted RPE cells will encounter in situ. Approximately 3146 cultured fetal RPE cells mm(-2) were seeded onto these three surfaces and grown in organ culture for 1, 7, or 14 days. Explants were bisected and examined histologically or analyzed with a scanning electron microscope. RPE nuclear density was measured on stained sections. Morphology and cell density were compared to cells seeded onto bovine corneal endothelial cell-extracellular matrix (BCE-ECM). In situ submacular RPE nuclear density was also measured in tissue sections of donor eyes ranging from 18 weeks gestation to 88 years of age to determine the effect of age on RPE density. Compared to cells seeded onto BCE-ECM at similar density, RPE cell coverage and cellular morphology on aged submacular human Bruch's membrane was poor at all time points. In contrast to cells on BCE-ECM, RPE cell density on Bruch's membrane decreased with time. In general, cell morphology on all three Bruch's membrane surfaces worsened by day-7 compared to day-L Although some cells were more pigmented on RPE basement membrane and the deep inner collagenous layer at day-7, poor cellular morphology indicated the remaining cells were not well differentiated. At day-14, the cells were uniform and cuboidal on BCE-ECM, with cell density similar to that at day-7 and similar to in situ density of young donors (