Method for screening material suppressing fine dust-induced retinal cell damage and composition for suppressing fine dust-induced retinal cell damage
A technology of retinal cells and screening methods, applied in the field of retinal cell damage compositions, to achieve the effect of inhibiting damage
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[0064] Preparation and immunofluorescent staining of micropigmented retinal cells
[0065] In a 5% (v / v) CO incubator, at a temperature of 37°C, 10% (v / v) of fetal bovine serum (FBS) and 1% of penicillin-streptomycin (Invitrogen) were added. Retinal pigment epithelial cells (human telomerase-immortalized retinal pigmented epithelial cells, htRPE cells) purchased from ATCC (American Type Culture Collection) were cultured in DMEM (Dulbecco's modified Eagle`s medium) in Carlsbad, California, USA.
[0066] After the human epidermal normal keratinocytes were transferred to Lab-TexTM 2-well chamber slides (Tnermo Scientific Company, Waltham, Massachusetts, U.S.A.) at a cell number of 0.5×105 per well, the cultured retinal pigment epithelial cells were Inoculate (6x104 cell number / well) on a 12-well plate, and after 24 hours, use dust PM10 (ERM-CZ100, sample number: 0504) purchased from ERM (European Reference Materials) at 100 μg / ml and 300 μg / ml respectively concentration, then ri...
experiment Embodiment 1
[0083] [Experimental Example 1] Recovery of retinal pigment epithelial cells caused by fine dust by formation of primary cilia
[0084] The primary cilium is an intracellular organ with a bipolar structure, and the combination (formation of cilia) and disassembly of cilia are interconnected through various causes such as cell cycle, polarity, vertebral development, and genetic diseases. It is known that primary cilia play an important role in the function of retinal pigmented epithelial cells. Specifically, retinal pigment epithelial cells play an important role in absorbing light and transporting substances in the eye. In order to complete this function well, good development of primary cilia is required. Therefore, changes in the expression of primary cilia when the retinal pigment epithelial cells were treated with fine dust were experimentally confirmed.
[0085] RPE cells were cultured in serum-free medium to confirm whether serum starvation induces primary cilia formati...
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