Comparison of in vitro isolation and culture of retinal pigment epithelial cells in rabbit eye and pig eye

Background Culture of retinal pigment epithelium(RPE) cells is very important for establishment of proliferative vitreoretinopathy (PVR) model,prevention and treatment of PVR as well as RPE cell transplantation.Isolation of animal RPE cells by trypsinization is a critical step.ObjectiveThe present study is to establish the methods of isolation and culture of retinal pigment epithelium (RPE) cells in rabbit and comparied with that of pig RPE culture.MethodsRPE cells were isolated by trypsinization in pigmented rabbit and pig and cultured in DMEM containing 20% fetal bovine serum.Cultured RPE cells were identified by immunochemistry.The fourth generations of cells were used in this experiment.Morphology and characteristics of cultured RPE cells from rabbit and pig were examined and compared under the light microscope.ResultsIsolated RPE cells from pig were obtained by once trypsin digestion,but two times of trypsinization were needed in rabbit RPE cells isolation.The differentiation in response to trypsinization was related to anatomic difference between the two types of cells .The adherence time of pig RPE cells was 24 hours ,however,the rabbit RPE needed 48-72 hours after culture.Proliferation and vitality of cultured cells were gradually attenuated and melanin decreased after several times subculture.The morphology of culture RPE cells was obviously different between rabbit and pig because species difference.Immunohistochemistry demonstrated the positive response of RPE cells for keratin.ConclusionRPE cells can be acquired from both rabbit and pig by trypsinization and culture.The culture process of RPE cells of pig is simpler than that of rabbit.Cells within the fourth generations are suitable for experimental application. Key words: retinal pigment epithelium cells; rabbit; pig; cell culture