Heat and protease treatment of aqueous humor: effect on cell DNA synthesis and growth.
Rabbit skin and Tenon's capsule fibroblasts were cultured in media containing 20% rabbit aqueous humor as a stimulatory agent. Cell stimulation was quantitated by 3H-thymidine uptake. The aqueous humor was either: (1) untreated; (2) heated at 37 degrees C for 60 min; (3) boiled for 2 min; or (4) heated at 37 degrees 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 degrees 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.
D000818:Animals; D001082:Aqueous Humor / Q000187:drug effects / Q000502:physiology*; D002452:Cell Count; D002478:Cells, Cultured; D003228:Conjunctiva / Q000378:metabolism*; D004247:DNA / Q000096:biosynthesis*; D005347:Fibroblasts / Q000378:metabolism; D006358:Hot Temperature; D011817:Rabbits; D012867:Skin / Q000378:metabolism*; D014357:Trypsin / Q000494:pharmacology*
D004247:DNA; D014357:Trypsin