Isolation and characterization of a spontaneously immortalized bovine retinal pigmented epithelial cell line.
Isolation and characterization of a spontaneously immortalized bovine retinal pigmented epithelial cell line.
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DOI：
10.1186/1471-2121-10-33
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发表时间：
2009-05-04
期刊：
影响因子：
--
通讯作者：
Gaillard ER
中科院分区：
文献类型：
--
作者：
Liggett TE;Griffiths TD;Gaillard ER
视网膜色素上皮(RPE)与眼睛的光感受器外节并列。光感受器细胞的接近是它们生存的先决条件，因为它们依赖RPE去除外节，也受到RPE细胞旁分泌因素的影响。RPE细胞死亡会导致光感受器功能的进行性丧失，这会降低视力，随着时间的推移，会导致失明。视网膜变性可导致多种视网膜病变，如Stargardt病、视锥细胞营养不良(CRD)、视网膜色素变性(RP)、黄斑眼底病(FFM)、Best病和老年性黄斑变性(AMD)。我们培养了原代牛RPE细胞，以进一步了解RPE细胞死亡的机制。其中一种名为tRPE的培养物超越了衰老，并进一步确定了其作为视网膜疾病模型的可行性。该tRPE细胞系已传代至150个群体倍增，并被证明在形态上与原代细胞相似。利用RPE特异基因RPE65和CRALBP以及RPE特异蛋白RPE65和Bestrophin，通过逆转录聚合酶链式反应和免疫细胞化学鉴定它们是RPE起源的。TRPE细胞对波形蛋白、细胞角蛋白和闭锁带-1抗体也呈免疫反应。染色体分析显示为正常的二倍体数量。TRPE细胞不能在悬浮或软琼脂中生长。在~3H胸腺嘧啶核苷掺入后，细胞融合后似乎没有明显的分裂。TRPE细胞是永生的，但仍表现出接触抑制、血清依赖性、单层生长和分泌细胞外基质。它们保留了体内的形态、基因表达和细胞极性。此外，细胞内吞外源性黑素、A2E和纯化的脂褐素颗粒。该细胞系可能是视网膜黄斑病变的一个有用的体外研究模型。
The Retinal Pigmented Epithelium (RPE) is juxtaposed with the photoreceptor outer segments of the eye. The proximity of the photoreceptor cells is a prerequisite for their survival, as they depend on the RPE to remove the outer segments and are also influenced by RPE cell paracrine factors. RPE cell death can cause a progressive loss of photoreceptor function, which can diminish vision and, over time, blindness ensues. Degeneration of the retina has been shown to induce a variety of retinopathies, such as Stargardt's disease, Cone-Rod Dystrophy (CRD), Retinitis Pigmentosa (RP), Fundus Flavimaculatus (FFM), Best's disease and Age-related Macular Degeneration (AMD). We have cultured primary bovine RPE cells to gain a further understanding of the mechanisms of RPE cell death. One of the cultures, named tRPE, surpassed senescence and was further characterized to determine its viability as a model for retinal diseases. The tRPE cell line has been passaged up to 150 population doublings and was shown to be morphologically similar to primary cells. They have been characterized to be of RPE origin by reverse transcriptase PCR and immunocytochemistry using the RPE-specific genes RPE65 and CRALBP and RPE-specific proteins RPE65 and Bestrophin. The tRPE cells are also immunoreactive to vimentin, cytokeratin and zonula occludens-1 antibodies. Chromosome analysis indicates a normal diploid number. The tRPE cells do not grow in suspension or in soft agar. After 3H thymidine incorporation, the cells do not appear to divide appreciably after confluency. The tRPE cells are immortal, but still exhibit contact inhibition, serum dependence, monolayer growth and secrete an extra-cellular matrix. They retain the in-vivo morphology, gene expression and cell polarity. Additionally, the cells endocytose exogenous melanin, A2E and purified lipofuscin granules. This cell line may be a useful in-vitro research model for retinal maculopathies.
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DOI：
10.1016/s0022-5320(66)80008-4
发表时间：
1966-01-01
期刊：
JOURNAL OF ULTRASTRUCTURE RESEARCH
影响因子：
--
作者：
BREATHNA.AS;WYLLIE, LMA
通讯作者：
WYLLIE, LMA
紧密连接链内和之间异质claudin物种相互作用的方式。
影响因子：
7.8
作者：
Furuse, M;Sasaki, H;Tsukita, S
通讯作者：
Tsukita, S
影响因子：
2
作者：
DUTT, K;WALDREP, JC;VERLY, G
通讯作者：
VERLY, G
DOI：
10.1083/jcb.97.3.703
发表时间：
1983-09
期刊：
The Journal of cell biology
影响因子：
--
作者：
Bunt-Milam AH;Saari JC
通讯作者：
Saari JC
DOI：
10.1083/jcb.127.6.1617
发表时间：
1994-12
期刊：
The Journal of cell biology
影响因子：
--
作者：
Furuse M;Itoh M;Hirase T;Nagafuchi A;Yonemura S;Tsukita S;Tsukita S
通讯作者：
Tsukita S