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ESTABLISHMENT OF HUMAN LENS EPITHELIAL CELL LINES

人晶状体上皮细胞系的建立

基本信息

批准号：
6518691
负责人：
John R. Reddan
金额：
$ 21.3万
依托单位：
OAKLAND UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2000
资助国家：
美国
起止时间：
2000-07-01 至 2005-05-31
项目状态：
已结题

项目摘要

The aim of cataract research is to prevent or delay human cataract formation. Cataract is the world's leading cause of blindness. Impaired epithelial function invariably leads to a loss of transparency. A therapy aimed at delaying or preventing human cataracts needs to be evaluated on human lens epithelial cells (HLECs). Due to the scarcity of viable human lenses, numerous investigators have tried to culture HLECs. However, normal, HLECs complete 2-4 passages in culture and undergo senescence. HLECs have properties different from those of the lower species. For example, lens epithelial lines can be established from lower species using standard techniques; this is not true of HLECs. Metabolic fluxes in various species are also different relative to the polyol pathway. Aldose reductase activity is high in the dog and rat lens, very low in the human lens and absent in the mouse. Although viral transformation of HLECs permits growth, virally transformed cells have properties that are different from their non-transformed counterparts. Experiments are needed to determine the properties of normal HLECs. Our aims are to establish a reliable supply of normal HLECs, characterize them and make them available to the eye research community. This is absolutely necessary for future studies. The aims are to: 1. Determine if a wound-healing environment permits the long-term growth of normal HLECs from young, old and cataractous lenses. 2. Determine if the introduction of telomerase into HLECs permits establishment of lines fromlens explants, permits established lines to overcome senescence, and if the presence of telomerase maintains the diploid state and allows maintenance of in vivo characteristics. Determine the ploidy, isoenzyme phenotyping, DNA fingerprint, alphaA and alphaB crystallin expression, and growth in cells generated Aim 1 or 2. 3. Determine if there are metabolic differences between cells from young, old and cataractous lenses. 4. Identify lens-specific regulatory elements in the promoters of genes that encode gamma-glutamylcysteine synthetase (GCS).

白内障研究的目的是预防或延缓人类白内障的形成。白内障是世界上致盲的主要原因。上皮功能受损总是导致透明性的丧失。一种旨在延缓或预防人类白内障的治疗方法需要在人晶状体上皮细胞（HLECs）上进行评估。由于缺乏可行的人类晶状体，许多研究者试图培养高晶状体。然而，正常的HLECs在培养中完成2-4代并经历衰老。HLECs具有不同于低等物种的特性。例如，可以使用标准技术从较低物种建立晶状体上皮细胞系；这对HLECs来说不是真的。不同物种的代谢通量也不同于多元醇途径。醛糖还原酶活性在狗和大鼠晶状体中很高，在人晶状体中很低，在小鼠中不存在。虽然病毒转化的HLECs允许生长，但病毒转化的细胞具有不同于非转化细胞的特性。需要通过实验来确定正常的HLECs的性质。我们的目标是建立正常hlec的可靠供应，对其进行表征，并将其提供给眼科研究界。这对今后的研究是绝对必要的。其目的是：1。确定伤口愈合的环境是否允许年轻、年老和白内障晶状体的正常HLECs长期生长。2. 确定端粒酶引入HLECs是否允许从外植体建立系，是否允许建立系克服衰老，以及端粒酶的存在是否维持二倍体状态并允许维持体内特征。测定其倍性、同工酶表型、DNA指纹图谱、α a和α b结晶蛋白表达和生长情况。3. 确定年轻、年老和白内障晶状体细胞之间是否存在代谢差异。4. 在编码γ -谷氨酰半胱氨酸合成酶（GCS）的基因启动子中鉴定晶状体特异性调控元件。