Cell density and solvent are critical parameters affecting formazan evaluation in MTT assay



Título del documento: Cell density and solvent are critical parameters affecting formazan evaluation in MTT assay
Revista: Brazilian archives of biology and technology
Base de datos: PERIÓDICA
Número de sistema: 000373464
ISSN: 1516-8913
Autors: 1
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Institucions: 1Universidade de Sao Paulo, Faculdade de Odontologia de Bauru, Bauru, Sao Paulo. Brasil
Any:
Període: May-Jun
Volum: 57
Número: 3
Paginació: 381-385
País: Brasil
Idioma: Inglés
Tipo de documento: Artículo
Enfoque: Analítico, descriptivo
Resumen en inglés 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; 1.156x104; 3.125x104; 1.156x105 and 3.125x105 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 number of cells and solvents: HCl/SDS (0 to 0.13), EtOH/HAc (0 to 0.22), DMSO (0.76 to 1.31) and PropOH (0.66 to 1.04). The DMSO and PropOH groups presented the most appropriate protocols for NIH/3T3 fibroblasts cell viability, especially at the density of 1.156x104 cells/cm2
Disciplines Biología
Paraules clau: Biología celular,
Cultivo de células,
Fibroblastos,
Viabilidad de células,
Densidad celular,
Bioensayos
Keyword: Biology,
Cell biology,
Cell culture,
Fibroblasts,
Cell viability,
Cell density,
Bioassays
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