| Backgrounds Age-related macular degeneration (ARMD) is a common reason that it causes blindness in old people in developed countries and regions, especially for the blinded population aged above fifty years in European countries and U.S., and the incidence of ARMD is higher with increase of age. Epidemiological investigation showed that the incidence of this disease in China is almost the same as that in other countries. Accoding to its clinic symptoms, ARMD is classified into dry and wet types and at the early stage it shows that retinal pigment epithelium (RPE) cells, especially macular RPE cell, are abnormal in cellular metabolism and bio-functions under induction by many factors, leading to the Drusen formation because of lipid deposition or the appearance of choroidal neovascularization (CNV) that further causes dry or wet ARMD and damages severely and irreversibly central vision as a result.Retinal pigment epithelium (RPE) cells are located at the outmost part of the 10 layers-composed retina which is constituted by successive monolayer of the hexagonal cells with the same size and regular morphology. The inner part of the pigment retina is connected with the photoreceptor outer segments of neural retina and disposes of shed photoreceptor outer segments by phagocytosis, playing a crucial role in renovation of the outer segments of ocular cells and maintenance of normal ocular functions. The morbidity in phagocytosis of the RPE will give rise to accumulation of rod outer segments (ROS) of the ocular cells and they undergo apoptosis. On the other hand, the RPE cells also form a blood-retinal barrier. Many RPE-related diseases are associated with visional functions-disordered eye diseases and choroidal deterioration.Objectives This study is based on the pathogeny of ARMD to analyze the culturing characteristics and phagocytosis of the RPE from human embryo, and to further discuss protective effects of LCVS1001 on UV-induced damaged RPE cells from human embryo and pathological changes at the molecular and cellular levels during the morbidity of ARMD, providing bases on exploiting specific, less-damage and effective treatment ways.Methods Fresh human embryo eyeballs were collected and after microdissection RPE cells were separated by mechanical and trypsin digestion. Using immunofluorescence, we investigated their phagocytosis to retinal outer segments. A UV-induced damaged model was established and protective effects of LCVS1001 on the RPE cells were evaluated through flow cytometry and RT-PCR.Results After the human embryo developed for 13-15 weeks, the RPE cells could be cultured in vitro according to standard culturing methods. Based on their growth curve, the RPE cells in generations 2-6 were collected for essays and they showed ability of phagocytosis. Intervening experiments indicated that LCVS1001 had protective effects on UV-induced damaged human embryo RPE cells and its action pathway is connected with p53.Conclusion 1. The culturing method of human embryo RPE cells is improved; 2. phagocytosis ability of human embryo RPE cells is proved in vitro; 3. a UV-induced damaged model is established ; 4.The protective effects on UV-induced damaged human embryo RPE cells are certified, which provides basis on farther studies of pathogeny and development of drugs against ARMD.
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