An improved method for isolation and culture of pigment epithelial cells from rat retina

An improved method for isolation and culture of pigment epithelial cells from normal rat retinas is described. Following a brief incubation in the neutral protease Dispase, large epithelial sheets can be harvested rapidly. The separation of the choroid from the pigment epithelium prior to retinal detachment greatly reduces the risk of contamination of the cultures with choroidal cells. Growth of pigment epithelial cells on Matrigel-coated microporous filters in hormonally-defined medium allows for development of high levels of transepithelial electrical resistance as well as for preservation of the differentiated pigment epithelial cell phenotype. This method should be useful for studies of pigment epithelial cell permeability and structural differentiation in vitro.