Objective To investigate the effect of peritoneal exudative cells as feeder cells on growth state of primary culture of adult rat retinal M uuml;ller cells. Methods Peritoneal exudative cells were gained from adult rats, which were identified with specifically biological marker of macrophage (CD68). The phagocytosis was evaluated by the ink particles experiment. Retinal M uuml;ller cells of adult rats were cultured by enzyme digestion method, and identified by GFAP and vimentin immunocytochemically. As the feeder cells, peritoneal exudative cells were cocultured with M uuml;ller cells. The proliferation cycle of M uuml;ller cells was assayed by flow cytometry. One-step TUNEL staining was employed to detect the apoptotic M uuml;ller cells. Results Over ninety-five percent of rat peritoneal exudative cells were macrophage, which have a favourable phagocytic ability for the ink particles. The primary cultured M uuml;ller cells adhered to the wall of flask and grew fast, with large applanate cell bodies. The third-generation cells grew slowly. After cocultured with feeder cells, the M uuml;ller cells showed more rapid growth rate with more cells in S and G2/M phase(S phase, t=4.172, P lt;0.001; G2/M phase, t=3.562, P lt;0.01） and less apoptotic rate (t=3.804, P lt;0.01). The growing cycle was cut down from 25-30 days to 1822 days for the firstgeneration cells, from 10-15 days to 7-10 days for the second-generation cells. Conclusion It is an effective method to use the peritoneal exudative cells as feeder cells cocultured with primary culture of retinal M uuml;ller cells, which can shorten the culture period of M uuml;ller cells in adult rats.
Citation： 毛俊峰,刘双珍,秦文娟,李凤云,谭浅,吴小影. Primary culture of retinal Müller cells in adult rats assisted by the peritoneal exudative cells. Chinese Journal of Ocular Fundus Diseases, 2009, 25(3): 206-209. doi: Copy