Les cellules sont ensemencées dans des plaques à 96 puits à une
densité de 1,5 10^ à 1,5 lO"^ cellules par puits et induites ou non par la
déxaméthasone 10'^ M. Journellement 20 ul d'une solution de MTT 5
mg/ml (3-(4, 5-diméthylthia2ol-2-yl)2, 5-diphényl tétrazolium bromide;
sigma) sont ajoutés aux 200 ul de milieu de culture. Une incubation de 2h.
permet le métabolisme du produit par la cellule. Le milieu est ensuite
enlevé et 200 ul de DMSO (diméthyl sulfoxide) sont ajoutés à chaque
puits pour solubiliser le MTT cristalisé dans les cellules.
La densité optique mesurée à 540 nm rend compte du nombre de
cellules en présence. Les courbes de croissance sont tracées en utilisant
les dilutions de cellules qui donnent des densités optiques non saturées
au cours des dix jours de culture.
82
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