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Cell density and solvent are critical parameters affecting formazan evaluation in MTT assay BABT
Gasque,Kellen Cristina da Silva; Al-Ahj,Luana Polioni; Oliveira,Rodrigo Cardoso; Magalhães,Ana Carolina.
The aim of this study was to establish the more accurate protocol for fibroblast cell viability using MTT assay. NIH/3T3 fibroblasts were seeded at the following cell densities: 3.125x10³; 1.156x10(4); 3.125x10(4); 1.156x10(5) and 3.125x10(5) cells/cm². Following 24h of seeding, MTT was added to the wells. After 4h of the MTT addition, different solvents were added to solubilize the formazan crystals: 1) HCl/SDS group- 20% SDS and 0.01 M HCl; 2) EtOH/ HAc group-50% ethanol and 1% acetic acid; 3) DMSO group- 99.5% dimethyl sulfoxide; and 4) PropOH group- 99.5% isopropanol. The absorbance values were measured using a spectrophotometer at 570 nm. The data were analyzed by 2-way ANOVA (p<0.05) and showed that the absorbance average varied according to the...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Cell viability; MTT assay; Fibroblast NIH/3T3; Solvents.
Ano: 2014 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132014000300011
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