THE EFFECTS OF GROWTH-FACTORS ON TENONS CAPSULE FIBROBLASTS IN SERUM-FREE CULTURE

This study was performed to develop and improve a completely defined i n vitro ocular wound-healing model of fibroblast proliferation for gla ucoma filtration surgery. This model is essential for the investigatio n of protein-sensitive drugs and cytokines. Tenon's capsule fibroblast s in their third passage were incubated overnight, washed free of seru m, and fed defined media, Aim V or Clonetics FBM serum-free medium con taining platelet-derived growth factor, basic fibroblast growth factor , epidermal growth factor, or fibronectin at various dilutions and in combinations at optimum concentrations. Proliferation was measured by H-3-thymidine incorporation at 1, 3, and 7 days. Morphology was compar ed to controls fed Minimum Essential Medium + 10% serum. Single factor s stimulated the greatest amount of thymidine uptake on day 3. Optimum concentrations were epidermal growth factor at 5 ng/ml, basic fibrobl ast growth factor at 10 ng/ml and platelet-derived growth factor at 20 ng/ml. Identical combinations of factors stimulated nearly twice the thymidine uptake in Clonetics medium as in Aim V. Epidermal growth fac tor activity was inhibited by either basic fibroblast growth factor or platelet-derived growth factor. Basic fibroblast growth factor and pl atelet-derived growth factor together produced a less than additive ef fect. The performance of either serum-free medium may be improved by t he addition of basic fibroblast growth factor or platelet-derived grow th factor. The optimum serum-free medium (Clonetics FBM) with growth f actors was unable to stimulate proliferation as much as Minimum Essent ial Medium + 10% NBS, but was successful in maintaining viability duri ng the 7 day test period.