Sparrow Janet R, Kim So Ra, Wu Yalin
Departments of Ophthalmology and Pathology and Cell Biology, Columbia University, New York, NY 10032, USA.
Methods Mol Biol. 2010;652:315-27. doi: 10.1007/978-1-60327-325-1_18.
Bisretinoid lipofuscin compounds that accumulate in retinal pigment epithelial (RPE) cells are implicated in the pathogenesis of some forms of macular degeneration. In the development of approaches to the amelioration of retinal disorders characterized by enhanced RPE lipofuscin formation, attention is being given to therapies that reduce the production of these damaging pigments. An understanding of the biosynthetic pathways by which these molecules form is essential to the development of these therapies. Thus methods for studying the biosynthesis of these compounds are presented. A tissue culture model is also described whereby a human RPE cell line that is otherwise devoid of bisretinoid lipofuscin compounds is employed and synthesized A2E is delivered to the cells. This approach allows for a population of RPE cells that have accumulated the lipofuscin fluorophore A2E in addition to A2E-free cells.
在视网膜色素上皮（RPE）细胞中积累的双视黄醛脂褐素化合物与某些形式的黄斑变性的发病机制有关。在开发改善以RPE脂褐素形成增强为特征的视网膜疾病的方法时，人们将注意力集中在减少这些有害色素产生的疗法上。了解这些分子形成的生物合成途径对于开发这些疗法至关重要。因此，本文介绍了研究这些化合物生物合成的方法。还描述了一种组织培养模型，即使用原本不含双视黄醛脂褐素化合物的人RPE细胞系，并将合成的A2E递送至细胞。这种方法可以产生一群除了不含A2E的细胞外还积累了脂褐素荧光团A2E的RPE细胞。
