Improved sensitivity and decreased sample size in a cytotoxicity test for biomaterials: a modified colony microassay using a microplate and crystal violet staining.
Modified Eagle's minimum essential medium supplemented with 5% fetal calf serum was highly sensitive to cytotoxicity and formed large control colonies in the V79 colony assay. A highly sensitive cytotoxicity test was developed using 96-well microtiter plates. Test chemicals or extracts of polyurethane materials containing the same chemicals were added 24 h after inoculation of cell suspensions. The cells were fixed and stained with crystal violet after additional culture for 6 days (V79 cells) or 10 days (Balb/3T3 cells). In terms of sensitivity and rapid quantitative measurement, this modified colony microassay, using a low cell density in 96-well microplates, was superior to various cytotoxicity tests such as colony, growth inhibition, cytolethality, and agar diffusion assays.
Tsuchiya
T
T
National Institute of Health Sciences, Tokyo, Japan.
Ikarashi
Y
Y
Arai
T
T
Ohhashi
J
J
Nakamura
A
A
D000818:Animals; D001672:Biocompatible Materials / Q000633:toxicity*; D002455:Cell Division / Q000187:drug effects*; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D016895:Culture Media, Serum-Free; D004305:Dose-Response Relationship, Drug; D005840:Gentian Violet; D008768:Methylmethacrylates / Q000633:toxicity*; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000379:methods; D013329:Structure-Activity Relationship; D013859:Thiocarbamates / Q000494:pharmacology*
D001672:Biocompatible Materials; D016895:Culture Media, Serum-Free; D008768:Methylmethacrylates; D013859:Thiocarbamates; D005840:Gentian Violet