Improved Culture Method of Retinal Pigment Epithelial Cells and Functional-morphological Characteristics In Vitro

ABSTRACT

To study the isolation and purification and proliferation of the cell in cell culture system, and to develop an improved culture method by a modified cell isolation technique and modified culture medium. The RPE cells were cultured in 3 different mediums: type I(MEM medium with 20% FCS) type II(F-10 medium with 20% FCS) and type III(DMEM medium with 10% FCS, EGF, hydrocortisone, insulin, ethanolamine, phosphoethanolamine, chorea toxin, triiodotyronine, adenine, transferrin and BPE). We compared population doubling(P.D.), population doubling time(P.D.T), morphologic changes and phagocytic activity during a 7week period. Rapid proliferation and high purity of retinal pigment epithelial cells(RPE cells) showed in type III culture medium. Type III culture medium presented the best results in P.D., P.D.T. and cell purification. In type III culture medium, single RPE cells produced about 6 X 10(7) RPE cells in the 7week period and morphology and phagocytic activity were well maintained, when UV-B irradiation at RPE was used to produce melanin, it had no effect, but the RPE cell was inhibited by UV-B irradiation. This improved culture method for RPE cells will provide a good in-vitro model for the studies of biochemistry, cellular function of the RPE cell, as well as its clinical application in eye disease.