Effects of Miiller cells on growth of RPE cells in a co-culture system in hypoxia condition

Abstract

Background More and more evidences suggest that the interaction of retinal pigment epithelial （RPE） cells and Mtiller cells plays an important role in the progression of retinal diseases. Objective Present study was to investigate the role of Mtiller cells in growth of RPE cells with a co-culture system in vitro under the hypoxia condition. Methods Mtiller cells and RPE cells of rabbit were primarily cultured and digested using explant culture method and trypsas method. Cultured Mtiller cells and RPE cells were identified by glial fibrillary acidic protein（GFAP） staining,S-l00 staining and cytokine-18 staining respectively. CoC12 was added into DMEM to establish the hypoxia models of Muiller cells and RPE cells. The third generations of Mtiller cells and RPE cells were co-cultured under the normoxia and hypoxia using transwell chamber, and the proliferation and migration of cultured RPE cells were examined using MTT and compared with only RPE ceils cultured group at 3, 6, 24 and 48 hours. Results Over 90% of primarily cultured RPE cells showed the positive response for CK-18. Cultured Mtiller cells presented the positive response for GFAP and S-100. The proliferation and migration of RPE cells were significantly increased in hypoxia condition compared with normoxia condition （P〈0.01）. In hypoxia group, amount of proliferation and migration of RPE cells in co-culture group were higher than the only RPE cells cultured group （P〈0. 01 ）. Conclusion Hypoxia condition promote the proliferation and migration of RPE cells. Mailer cells appear to aggravate the proliferation and migration of RPE cells under the hypoxia status. Key words: Muller cell; Retinal pigment epithelium; Hypoxia; Cellular proliferation; Cellular migration