​上海易色医疗科技有限公司

Shanghai Yise Medical Technology Co.,Ltd.

杂志：Anticancer Res. 1999 Mar-Apr;19(2A):1245-8.

标题：Falsification of tetrazolium dye (MTT) based cytotoxicity assay results due to mycoplasma contamination of cell cultures.

作者：Denecke J1, Becker K, Jürgens H, Gross R, Wolff JE.

摘要：Mycoplasma contamination of cell cultures is a frequently observed problem. Due to the inconspicuous growth in cell cultures, periodical screening procedures represent the only protection. Many influences of mycoplasma on cell culture parameters have been described. We addressed the question of whether mycoplasma contamination affects the most frequently used cytotoxicity assay, the tetrazolium based MTT assay. We contaminated C6 glioma cells with mycoplasma and performed MTT assays with doxorubicin, vincristine, etoposide and cisplatinum under various conditions. Contaminated cells demonstrated significant different results when tested with the MTT assay than mycoplasma free controls. Differences were not detectable when cells were counted as toxicity assay. Due to an additional reduction of tetrazolium by mycoplasmas, contaminated cells appeared up to 15 fold resistant to doxorubicin, vincristine and etoposide, but not to cisplatinum. Differences decreased with decreasing drug doses and decreasing plated cell count. Our findings confirm the compelling need for periodical mycoplasma screening, especially when tetrazolium based cytotoxicity assay (MTT) are used.

主要结论: 由于支原体对MTT的额外还原作用，对阿霉素（Doxorubicin, 一种抗肿瘤药物）的抗性，支原体污染和非污染的细胞二者相差15倍.​可以想象由于细胞支原体的污染也极有可能导致CCK8实验出现类似的错误。