Sieuwerts A M, Klijn J G, Peters H A, Foekens J A
Division of Endocrine Oncology (Department of Medical Oncology), Dr. Daniel den Hoed Cancer Center, Rotterdam, The Netherlands.
Eur J Clin Chem Clin Biochem. 1995 Nov;33(11):813-23. doi: 10.1515/cclm.1995.33.11.813.
The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide (MTT) assay is widely used for in vitro measurement of the metabolic viability of cell cultures subjected to different culture conditions. This convenient assay, which is based on the ability of viable cells to produce formazan, can be affected significantly by a number of conditions. These conditions can be roughly divided into groups, firstly influences which affect the spectrum of the produced formazan and secondly influences which affect the amount of formazan produced per cell. We studied the various chemical and biochemical aspects involved in the MTT assay. Our data indicated that microscopical viewing of cell cultures before and after performing the assay, a medium renewal with a well-defined MTT-incubation medium at the end of the culture period and regular cell counting are essential steps to ensure a reliable performance of the MTT assay. In conclusion, providing the necessary precautions are taken, the MTT assay can be used reliably to measure metabolic activity of cell cultures in vitro for the assessment of growth characteristics, IC50-values and cell survival.
3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）法被广泛用于体外测量处于不同培养条件下的细胞培养物的代谢活力。这种便捷的检测方法基于活细胞产生甲臜的能力，会受到多种条件的显著影响。这些条件大致可分为两类，第一类是影响所产生甲臜光谱的因素，第二类是影响每个细胞产生甲臜量的因素。我们研究了MTT法涉及的各种化学和生化方面。我们的数据表明，在进行检测前后对细胞培养物进行显微镜观察、在培养期结束时用明确的MTT孵育培养基更换培养基以及定期进行细胞计数是确保MTT法可靠运行的关键步骤。总之，只要采取必要的预防措施，MTT法可可靠地用于体外测量细胞培养物的代谢活性，以评估生长特性、IC50值和细胞存活率。
