Isolation of a retinal pigment epithelial cell-derived fraction which promotes Müller cell proliferation

Retinal pigment epithelial cells (RPE) secrete a factor(s) in vitro which promotes Müller cell proliferation. To begin the isolation of the active factor, medium conditioned by cultured RPE (RPE-CM) was fractionated by fast protein liquid chromatography. The fractions were tested in an assay for Müller cell proliferation and it was found that one fraction contained biological activity comparable to that of complete RPE-CM. Gel electrophoresis demonstrated that this fraction consisted of proteins with approximate molecular masses of 35-88 kDa. An affinity assay revealed that only an 88-kDa protein in the RPE-CM binds to Müller cells. This protein was also present in the active fraction; therefore, it is the most likely source of the mitogenic activity of RPE-CM.