Rapid quantitative assays for corneal endothelial cell viability in vitro.
The Janus green photometric technique and the Mosmann's colorimetric MTT-assay to quantify corneal endothelial cytotoxicity were compared. Detergents, studied before by in vitro corneal perfusion, proved to be toxic at the 1% level in the Janus green assay with human corneas and at the 0.4% level in the MTT-assay with bovine endothelial cells. These results correlated well with earlier findings in vitro and in vivo. The use of human corneas reduced the ability of the Janus green photometric technique as a fast screening method because of the heterogeneity and restricted availability of this tissue. With the MTT-assay, large test series could be performed at the same time using cultured cells. Both assays are good tools to screen the cytotoxicity of chemicals and reduce the number of animals needed for endothelial cytotoxicity testing.
Pels
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E
Department of Morphology, Netherlands Ophthalmic Research Institute, Amsterdam.
Nuyts
R M
RM
Breebaart
A C
AC
Hartmann
C
C
D000818:Animals; D001391:Azo Compounds; D001681:Biological Assay; D002417:Cattle; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D004396:Coloring Agents; D003902:Detergents / Q000633:toxicity; D004728:Endothelium, Corneal / Q000166:cytology* / Q000187:drug effects; D006801:Humans; D010783:Photometry / Q000379:methods; D013778:Tetrazolium Salts; D013844:Thiazoles
D001391:Azo Compounds; D004396:Coloring Agents; D003902:Detergents; D013778:Tetrazolium Salts; D013844:Thiazoles; C047446:janus green; C022616:thiazolyl blue