van de Loosdrecht A A, Beelen R H, Ossenkoppele G J, Broekhoven M G, Langenhuijsen M M
Department of Hematology (BR238), University Hospital, Vrije Universiteit, Amsterdam, Netherlands.
J Immunol Methods. 1994 Sep 14;174(1-2):311-20. doi: 10.1016/0022-1759(94)90034-5.
The MTT-colorimetric monocyte mediated cytotoxicity assay, based upon the ability of living cells to reduce 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyltetrazolium bromide (MTT) into formazan, was evaluated using leukemic cells from five representative human leukemic cell lines and from 28 patients with acute myeloid leukemia (AML). An excellent linearity between absorbance and leukemic cell number was observed up to 5 x 10(4) cells/well and 50 x 10(4) cells/well for all cell lines and patients samples tested, respectively, in a 96-wells microtiter culture system. A huge variability in the susceptibility of leukemic cells to purified and IFN-gamma-activated human monocytes could be observed at effector-to-target cell (E:T) ratios of 1. The mean signal-to-noise ratio of the MTT assay for monocyte-leukemic cell mixtures from patients was 2.69 +/- 0.39 at E:T 1. In conclusion, the MTT based monocyte mediated cytotoxicity assay should be useful for studying the susceptibility of a variety of leukemic cells from cell lines and from patients with AML to monocytes in a rapid, sensitive and semi-automated manner.
基于活细胞将3-[4,5-二甲基噻唑-2-基]-2,5-二苯基溴化四氮唑（MTT）还原为甲臜的能力，采用来自5种代表性人类白血病细胞系以及28例急性髓性白血病（AML）患者的白血病细胞，对MTT比色法单核细胞介导的细胞毒性试验进行了评估。在96孔微量培养系统中，对于所有测试的细胞系和患者样本，分别在每孔5×10⁴个细胞和每孔50×10⁴个细胞时，观察到吸光度与白血病细胞数量之间具有良好的线性关系。在效应细胞与靶细胞（E:T）比例为1时，可观察到白血病细胞对纯化的和经γ干扰素激活的人单核细胞的敏感性存在巨大差异。在E:T为1时，患者单核细胞-白血病细胞混合物的MTT试验平均信噪比为2.69±0.39。总之，基于MTT的单核细胞介导的细胞毒性试验对于快速、灵敏且半自动地研究来自细胞系和AML患者的多种白血病细胞对单核细胞的敏感性应是有用的。
