Zhang Z D, Cox G
Father Sean O'Sullian Research Center, St. Joseph's Hospital, Hamilton, Ontario, Canada.
Biochem Mol Biol Int. 1996 Mar;38(3):431-6.
In this study, we evaluated the accuracy of the MTT (a tetrazolium salt) assay for cell counting by comparing it to haemocytometer. When airway smooth muscle cells were cultured in RPMI with fetal calf serum < or = 5%, no significant differences were observed in cell number counted by the two methods. For cells cultured in 10% serum, however, the result was 20% higher when counted by MTT assay. Rhodamine 123 uptake was similar in ASMCs cultured in the presence of serum at 5 and 10% indicating no difference in total mitochondrial activity per cell. Serum at > 5% may modulate the activity of enzymes responsible for reduction of MTT. Our results indicate that the MTT assay may not be accurate under certain conditions, especially when the treatment of cells can affect the enzymes that account for MTT reduction in addition to stimulating cell growth.
在本研究中，我们通过将MTT（一种四唑盐）分析法与血细胞计数器进行比较，评估了其用于细胞计数的准确性。当气道平滑肌细胞在含有小于或等于5%胎牛血清的RPMI培养基中培养时，两种方法所计数的细胞数量没有显著差异。然而，对于在10%血清中培养的细胞，用MTT分析法计数时结果高出20%。在5%和10%血清存在的情况下培养的气道平滑肌细胞中罗丹明123摄取情况相似，表明每个细胞的总线粒体活性没有差异。血清浓度大于5%可能会调节负责MTT还原的酶的活性。我们的结果表明，MTT分析法在某些条件下可能不准确，特别是当细胞处理除了刺激细胞生长外还能影响负责MTT还原的酶时。
