Health & Environmental Research Online (HERO)


Print Feedback Export to File
2982219 
Journal Article 
An improved colorimetric assay for cell proliferation and viability utilizing the tetrazolium salt XTT 
Roehm, NW; Rodgers, GH; Hatfield, SM; Glasebrook, AL 
1991 
Yes 
Journal of Immunological Methods
ISSN: 0022-1759 
142 
2 
257-265 
English 
1919029 
WOS:A1991GG60500015 
A new tetrazolium salt XTT, sodium 3'-[1-[(phenylamino)-carbonyl]-3,4-tetrazolium]-bis(4-methoxy-6- nitro)benzene-sulfonic acid hydrate, was evaluated for use in a colorimetric assay for cell viability and proliferation by normal activated T cells and several cytokine dependent cell lines. Cleavage of XTT by dehydrogenase enzymes of metabolically active cells yields a highly colored formazan product which is water soluble. This feature obviates the need for formazan crystal solubilization prior to absorbance measurements, as required when using other tetrazolium salts such as MTT. Bioreduction of XTT by all the murine cells examined was not particularly efficient, but could be potentiated by addition of electron coupling agents such as phenazine methosulfate (PMS) or menadione (MEN). Optimal concentrations of PMS or MEN were determined for the metabolism of XTT by the T cell lines HT-2 and 11.6, NFS-60 a myeloid leukemia, MC/9 a mast cell line and mitogen activated splenic T cells. When used in combination with PMS, each of these cells generated higher formazan absorbance values with XTT than were observed with MTT. Thus the use of XTT in colorimetric proliferation assays offer significant advantages over MTT, resulting from reduced assay time and sample handling, while offering equivalent sensitivity. 
Coloring Agents; Interleukin-2; Tetrazolium Salts; Thiazoles; Concanavalin A; 11028-71-0; 2,3-bis(2-methoxy-4-nitro-5-sulfophenyl)-5-((phenylamino)carbonyl)-2H-tetrazolium hydroxide; 117038-70-7; Vitamin K; 12001-79-5; Interleukin-4; 207137-56-2; Methylphenazonium Methosulfate; 299-11-6; thiazolyl blue; EUY85H477I; Index Medicus; Cell Survival -- drug effects; Interleukin-2 -- pharmacology; Spleen -- cytology; Dose-Response Relationship, Drug; Animals; Cell Division -- drug effects; Interleukin-4 -- pharmacology; Leukemia, Myeloid -- pathology; Mast Cells -- cytology; In Vitro Techniques; T-Lymphocytes -- cytology; Colorimetry -- methods