Simple colorimetric cell-cell adhesion assay using MTT-stained leukemia cells.
A new and simple method for quantitating adhesion of leukemia cells, HL60, to endothelial cells was developed. HL60 cells were incubated with MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) which forms a blue dye of formazan in the cells. MTT did not inhibit shape change of HL60 cells induced on activated endothelial cells at 90 min. The expression of a cell adhesion molecule, LFA-1, in HL60 cells at 21 h was not inhibited by MTT. After incubation of the MTT-labeled cells and endothelial cells, non-adhered cells were washed out. Adhered cells were lysed with dimethylsulfoxide, and quantitated by measuring absorbance at 540 nm. The absorbance was well correlated with the adherent cell numbers measured by direct counting or by 51Cr-labeling method. U937 and Ramos cells were also quantitatively labeled by MTT. The method has the advantage of being easy, simple and applicable to various cell-cell adhesion assays.
Pharmaceutical Research Laboratories, Kyowa Hakko Kogyo Co., Ltd., Shizuoka, Japan.
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I
Miki
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Ishihara
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Otoshi
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H
Kase
D002448:Cell Adhesion; D003124:Colorimetry; D004730:Endothelium, Vascular / Q000166:cytology*; D006801:Humans; D066298:In Vitro Techniques; D016169:Lymphocyte Function-Associated Antigen-1 / Q000378:metabolism; D010084:Oxidation-Reduction; D013778:Tetrazolium Salts; D014407:Tumor Cells, Cultured
D016169:Lymphocyte Function-Associated Antigen-1; D013778:Tetrazolium Salts