Culturing of Retinal Pigment Epithelial Cells on an Ex Vivo Model of Aged Human Bruch's Membrane.
Culturing of Retinal Pigment Epithelial Cells on an Ex Vivo Model of Aged Human Bruch's Membrane.
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DOI：
10.3791/57084
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发表时间：
2018-04-12
期刊：
影响因子：
--
通讯作者：
Fields MA
中科院分区：
文献类型：
--
作者：
Cai H;Gong J;Del Priore LV;Tezel TH;Fields MA
除了年龄相关眼病研究推荐的维生素和抗氧化剂外，对于占 90% 病例的“干性”或萎缩性年龄相关性黄斑变性 (AMD) 没有有效的治疗方法。需要通过治疗来减缓或延缓地理萎缩 (GA) 的发展，了解布鲁赫膜病理学是这一过程的一部分。人类布鲁赫膜的改变会导致视网膜色素上皮 (RPE) 细胞的损伤，从而导致 AMD 的进展。鉴于缺乏足够的动物模型来研究 AMD，老年人类布鲁赫膜的离体模型可作为研究永生化和原代细胞系以及诱导多能干细胞 (iPSC) 衍生的 RPE 系的 RPE 细胞行为的有用工具。在这里，我们提出了一种详细的方法，可以确定接种在从人类供体收获的人类布鲁赫膜外植体上的 RPE 细胞行为的影响，包括附着、凋亡和增殖、吞噬光感受器外节的能力、极性的建立和基因表达。该测定提供了老化 Bruch 膜的离体模型，以评估 RPE 细胞接种在老化/受损的细胞外基质上时的功能特征。
Aside from vitamins and antioxidants recommended by the Age-Related Eye Disease Study, there is no effective therapy for "dry," or atrophic age-related macular degeneration (AMD) which represents 90% of the cases. Therapies are needed to slow or retard the development of geographic atrophy (GA), and understanding Bruch's membrane pathology is part of this process. Alterations in human Bruch's membrane precede the progression of AMD by contributing to the damage of retinal pigment epithelial (RPE) cells. Given the lack of sufficient animal models to study AMD, ex vivo models of aged human Bruch's membrane serve as a useful tool to study the behavior of RPE cells from immortalized and primary cell lines as well as RPE lines derived from induced pluripotent stem cells (iPSCs). Here, we present a detailed method that allows one to determine the effects of RPE cell behavior seeded on harvested human Bruch's membrane explants from human donors, including attachment, apoptosis and proliferation, ability to phagocytize photoreceptor outer segments, establishment of polarity, and gene expression. This assay provides an ex vivo model of aged Bruch's membrane to assess the functional characteristics of RPE cells when seeded on aged/compromised extracellular matrix.
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DOI：
10.1097/opx.0000000000000322
发表时间：
2014-08
期刊：
Optometry and vision science : official publication of the American Academy of Optometry
影响因子：
--
作者：
Fletcher EL;Jobling AI;Greferath U;Mills SA;Waugh M;Ho T;de Iongh RU;Phipps JA;Vessey KA
通讯作者：
Vessey KA
影响因子：
3.9
作者：
Nguyen HV;Li Y;Tsang SH
通讯作者：
Tsang SH
影响因子：
3.4
作者：
Gullapalli, VK;Sugino, IK;Zarbin, MA
通讯作者：
Zarbin, MA
影响因子：
3.4
作者：
Castellarin, AA;Sugino, IK;Zarbin, MA
通讯作者：
Zarbin, MA
影响因子：
2
作者：
Cai, H;Del Priore, LV
通讯作者：
Del Priore, LV