HUMAN RETINAL CELL LINES--A MODEL FOR RETINAL DEVELOPMENT

人类正常视网膜发育和营养不良的机制是 仍然没有完全理解，尽管事实是超过一半的 每年失明的美国人被诊断出患有某种基因 叛逃。阐明正常视网膜发育中的事件可能会让我们 了解营养不良的基础及其更好的处理。多数 关于人类视网膜发育的可用信息的一部分来自 禽类啮齿动物模型和很少的人类研究局限于肿瘤细胞 台词。由于样本有限，目前还不可能进行长期研究 原代培养中的祖细胞和衰老。我们建议 一种替代战略，使我们能够进行长期研究。 这项提议的目标是建立年龄受限的细胞系 用SV-40T从发育中的(4、8、12个月胎儿)人视网膜组织 基因转染与近视眼视网膜发育的研究进展 同质的细胞群体。我们已经建立了一个视网膜细胞 Line(达特等人，1994,1996)。人们普遍认为视网膜细胞是多细胞的。 潜力，但这种能力取决于年龄吗？我们提出的关键问题 问：a)决定细胞命运的因素是固有的 祖细胞和发育变化，b)是细胞和分子 细胞命运特异性祖细胞微环境的变化 决定？我们建议检验细胞系产生的假设 来自不同年龄的人将受到发育限制，并将在 它们能够生成不同的细胞组合，这要么是由于1) 内源信号(谱系)的变化，2)外源信号的变化， 或3)改变了祖细胞对信号的反应能力 (外源性线索和内源性线索之间的相互作用)。在这项研究中，使用 细胞和分子方法，我们将确定谱系的作用 视网膜发育中的细胞外信号。我们建议 完成以下目标：1)建立年龄受限的细胞系 SV-40T基因转染法发育人视网膜。2)确定单元格 血统在不同年龄段的祖细胞中发挥的作用 通过内源信号。3)确定内生和外生的作用 信号以及它们如何在细胞命运决定中相互作用。

Mechanisms underlying normal human retinal development and dystrophies are still not fully understood, despite the fact that more than half of Americans who become blind each year are diagnosed with some genetic defect. Elucidation of events in normal retinal development may allow us to understand the basis of dystrophies and their better management. Most of the information available on human retinal development emanates from avian of rodent models and few human studies are limited to tumor cell lines. No long term studies are currently possible due to limited sampling of progenitors and senescence that ensues in primary cultures. We propose an alternative strategy that will enable us to perform long term studies. The objective of this proposal is to establish age-restricted cell lines from developing (4, 8, 12 months fetal) human retinal tissues by SV-40T gene transfection and elucidate events in nearly retinal development in homogenous cell populations. We have already established a retinal cell line (Dutt et al 1994, 1996). It is accepted that retinal cells are multi- potential but is this capacity age dependent? The key questions we propose to ask: a) are the factors that determine cell fate intrinsic to progenitors and change developmentally, b) are cellular and molecular changes in micro environment of progenitors specific for cell fate decisions? We propose to test the hypothesis that cell lines generated from different ages will be developmentally restricted and will vary in their ability to generate different combinations of cells either due to 1) changes in endogenous signals (lineages), 2) changes in exogenous signals, or 3) changed ability of the progenitors to respond to signals (interaction between exogenous and endogenous cues). In this study, using cellular and molecular approaches, we will determine the role of lineages versus extracellular signals in retinal development. We propose to accomplish the following aims: 1) Establish age restricted cell lines from human developing retina by SV-40T gene transfection. 2) Determine cell lineages in progenitors from different ages to determine the role played by endogenous signals. 3) Determine the role of endogenous and exogenous signals and how they interact in cell fate determination.