Heat and protease treatment of aqueous humor: effect on cell DNA synthesis and growth

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Abstract
Rabbit skin and Tenon's capsule fibroblasts were cultured in media containing 20% rabbit aqueous humor as a stimulatory agent. Cell stimulation was quantitated by3H-thymidine uptake. The aqueous humor was either: (1) untreated; (2) heated at 37° C for 60 min; (3) boiled for 2 min; or (4) heated at 37° C for 60 min with 0.025% trypsin, followed by boiling. The effects of the various treatments of aqueous humor on cell stimulation were compared among themselves and to 10% fetal bovine serum. It was found that: (1) heating of aqueous humor at 37° C for up to 60 min had no apparent effect on its ability to stimulate DNA synthesis in culture; (2) boiling and trypsin plus boiling reduced the ability of 20% aqueous humor to promote thymidine uptake to varying degrees; (3) 20% untreated aqueous humor was more effective than 10% fetal bovine serum in stimulating DNA synthesis in Tenon's capsule fibroblasts; skin fibroblasts responded similarly to both media.