Comparative analysis of three methods to assess viability of mammalian cells in culture.
Release of lactate dehydrogenase (LDH) from the cytoplasmic compartment, trypan blue exclusion and methylthiazole tetrazolium (MTT) colorimetric assays were compared with regard to their sensitivity in detecting damage of human cultured epithelial cells induced by sodium fluoride or puromycin. LDH assay did not detect any difference between controls and cells treated with either of the two drugs. Cell monolayers treated with 0.3% sodium fluoride or 10(-2) M puromycin presented higher percentages of cells that took up the trypan blue dye than controls but monolayers treated with lower drug concentrations did not differ from controls. Viability measured by MTT assay was the most sensitive assay, detecting a dose-dependent impairment of cell function after treatment with the two drugs. Moreover, MTT offered major advantages in speed, simplicity and precise quantitation over the other viability assays.
da Costa
A O
AO
Department of Microbiology and Immunology, State University of Rio de Janeiro UERJ, Brazil.
de Assis
M C
MC
Marques
E de A
Ede A
Plotkowski
M C
MC
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002470:Cell Survival; D002522:Chlorocebus aethiops; D004396:Coloring Agents; D006801:Humans; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008099:Liver / Q000166:cytology*; D008322:Mammals; D013778:Tetrazolium Salts; D013844:Thiazoles; D014343:Trypan Blue; D014709:Vero Cells
D004396:Coloring Agents; D013778:Tetrazolium Salts; D013844:Thiazoles; D007770:L-Lactate Dehydrogenase; C022616:thiazolyl blue; D014343:Trypan Blue