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

• Published in: Graefe's archive for clinical and experimental ophthalmology 1985, Vol.222 (3), p.154-157
• Main Authors: LITIN, B. S, JONES, M. A, HERSCHLER, J
• Format: Article
• Language: English
• Subjects: Animals; Aqueous Humor - drug effects; Aqueous Humor - physiology; Biological and medical sciences; Cell Count; Cells, Cultured; Conjunctiva - metabolism; DNA - biosynthesis; Eye and associated structures. Visual pathways and centers. Vision; Fibroblasts - metabolism; Fundamental and applied biological sciences. Psychology; Hot Temperature; Rabbits; Skin - metabolism; Trypsin - pharmacology; Vertebrates: nervous system and sense organs
• ISSN: 0721-832X; 1435-702X
• DOI: 10.1007/BF02173541

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.