Human serum reduces mitomycin-C cytotoxicity in human tenon's fibroblasts

Investigative Ophthalmology & Visual Science
Jonathan G CrowstonPaul R Healey

Abstract

To determine the effect of human serum factors on mitomycin-C (MMC) cytotoxicity in cultured human subconjunctival Tenon's capsule fibroblasts. Fibroblast monolayers were treated with 5-minute applications of mitomycin-C (0.4 mg/mL) and incubated in culture medium with or without additional human serum. Fibroblast apoptosis was quantified by direct cell counts based on nuclear morphology, flow cytometry with annexin-V/propidium iodide, and a lactate dehydrogenase release assay. The number of viable fibroblasts and fibroblast proliferation were measured with a colorimetric MTT assay and by bromodeoxyuridine (BrdU) labeling. Mitomycin-C induced significant levels of fibroblast apoptosis. The addition of human serum resulted in a 40% reduction in MMC-induced fibroblast apoptosis (range, 31.3%-55.3%; P = 0.021) as determined by nuclear morphology and a 32.4% reduction measured by annexin-V/PI. There was a corresponding dose-dependent increase in the number of viable fibroblasts. Serum did not restore proliferation in MMC-treated fibroblasts. Factors present in human serum reduce MMC cytotoxicity in cultured human Tenon's fibroblasts. Human serum increased the number of viable fibroblasts by inhibiting MMC-induced fibroblast apoptos...Continue Reading

Citations

Nov 10, 2009·Graefe's Archive for Clinical and Experimental Ophthalmology = Albrecht Von Graefes Archiv Für Klinische Und Experimentelle Ophthalmologie·Lars ChoritzHagen Thieme
Dec 4, 2010·Eye·P AgrawalV Sung
Sep 18, 2007·Journal of Glaucoma·Paul R HealeyWilliam Smiddy
Nov 12, 2010·Drug Metabolism and Disposition : the Biological Fate of Chemicals·Yongmei PanPeter W Swaan
Aug 18, 2016·Journal of Cataract and Refractive Surgery·Francisco Irochima PinheiroAcácio Alves de Souza Lima Filho
Nov 15, 2014·PloS One·Hermann AgisWilliam V Giannobile
Mar 21, 2012·Langmuir : the ACS Journal of Surfaces and Colloids·Aurora ColomerLourdes Pérez

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