Summary
The aim of this study was to optimize the experimental conditions of the MTT assay for primary cultures of human brain tumors. This assay is based on the mitochondrial reduction of MTT-(3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) salt to formazan crystals by living cells. Formazan can be quantified spectrophotometrically. This assay measures the antimetabolic and, by using an adequate recovery period for the cells, also the antiproliferative effects of cytotoxic drugs. Our results suggest the following experimental design for its application as an chemosensitivity assay for human brain tumors: 1-h drug exposure followed by a seven days recovery period without drugs. Then tumor cells are incubated 4 hours with 1 mg MTT/ml and final absorbance readings are performed at 550 nm and 630 nm as test and reference wavelengths respectively. In this way, the assay seems to be a reliable and simple method for rapid chemosensitivity testing in human brain tumors.
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References
Yung WKA: In vitro chemosensitivity testing and its clinical application in human gliomas. Neurosurg Rev 12: 197–203, 1989
Kimmel DW, Shapiro JR, Shapiro WR: In vitro drug sensitivity testing in human gliomas. J Neurosurg 66: 161–171, 1987
Von Hoff DD: He's not going to talk about in vitro predictive assays again, is he? J Nat Cancer Inst 82: 96–101, 1990
Mosmann T: Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65: 55–63, 1983
Altman FP: Tetrazolium salts and formazans. Prog Histochem Cytochem 9 (3): 1–56, 1976
Slater TF, Sawyer B, Sträuli U: Studies on succinate-tetrazolium reductase systems. Biochem Biophys Acta 77: 383–393, 1963
Carmichael J, DeGraff WG, Gazdar AF, Minna JD, Mitchell J: Evaluation of a tetrazolium-based semiautomated colorimetric assay: assessment of chemosensitivity testing. Cancer Res 47: 936–942, 1987
Cole SPC: Rapid chemosensitivity testing of human lung tumor cells using the MTT assay. Cancer Chemother Pharmacol 17: 259–263, 1986
Denizot F, Lang R: Rapid colorimetric assay for cell growth and survival. J Immunol Methods 89: 271–277, 1986
Ruben RL, Neubauer RH: Semiautomated colorimetric assay for in vitro screening of anticancer compounds. Cancer Treat Rep 71: 1141–1149, 1987
Twentyman PR, Luscombe M: A study of some variables in a tetrazolium dye (MTT) based assay for cell growth and chemosensitivity. Br J Cancer 56: 279–285, 1987
Nikkhah G, Darling JL, Thomas DGT: The application of the MTT chemosensitivity assay to short-term cell lines derived from gliomas: characterization and comparison with a 35S-methionine uptake assay. Reg Cancer Treat 2: 112–118, 1989
Nikkhah G, Torn JC, Hoffmann O, Kraemer HP, Darling JH, Schachenmayr W, Schönmayer R: The MTT-assay for chemosensitivity testing of human tumors of the central nervous system. Part II: Evaluation of patient- and drug-specific variables. J Neurooncol 13: 309–320
Carmichael J, DeGraff WG, Gazdar AF, Minna JD, Mitchell JB: Evaluation of a tetrazolium-based semiautomated colorimetric assay: assessment of radiosensitivity. Cancer Res 47: 943–946, 1987
Manthorpe M, Fagnini R, Skaper SD, Varon S: An automated colorimetric microassay for neurotrophic factors. Dev Brain Res 25: 191–198, 1986
Pieters R, Huismans DR, Leyva A, Veerman AJP: Comparison of the rapid automated MTT-assay with a dye exclusion assay for chemosensitivity testing in childhood leukaemia. Br J Cancer 59: 217–220, 1989
Twentyman PR, Fox NE, Rees JKH: Chemosensitivity testing of fresh leukaemia cells using the MTT colorimetric assay. Br J Haematol 71: 19–24, 1989
Shapiro JR, Shapiro WR: The subpopulations and isolated cell types of freshly resected high grade human gliomas: Their influence on the tumor's evolution in vivo and behaviour and therapy in vivo. Cancer Metastasis Rev 4: 107–124, 1985
Jabbar SAB, Twentyman PR, Watson JV: The MTT assay underestimates the growth inhibitory effects of interferons. Br J Cancer 60: 523–528, 1989
Bosanquet AG: Stability of solutions of antineoplastic agents during preparation and storage for in vitro assays. Cancer Chemother Pharmacol 17: 1–10, 1986
Bruhn HD: Zytostatika-Fibel: einschließich der wichtigsten Chemotherapie-Schemata, p. 3, Schattauer, New York, 1985
Bioscience Communications Inc.: Mode of action, 1–12. Novantrone (mitoxantrone) product monograph, Lederle International (ed), Lederle International, New York, 1988
Darling JL, Thomas DGT: Assay of anticancer drugs in tissue culture: the effect of cell kinetics and the mode of action of drugs on the chemosensitivity of cultures derived from malignant brain tumors. In: Chatel, Darcel and Pecker (eds) Brain Oncology, Martinus Nijhoff Publishers, Dordrecht, 1987, pp 369–371
Merry S, Kaye SB, Freshney RI: Cross-resistance to cytotoxic drugs in human glioma cell lines in culture. Br J Cancer 50: 831–835, 1984
Morgan D, Freshney RI, Darling JL, Thomas DGT, Celik F: Assay of anticancer drugs in tissue culture: cell cultures of biopsies from human astrocytoma. Br J Cancer 47: 205–214, 1983
Osieka R, Seeber S, Schmidt CG: Predictive tests in cancer chemotherapy: a reappraisal. Klin Wochenschr 62: 203–212, 1984
Thomas DGT, Darling JL, Freshney RI, Morgan D: In vitro chemosensitivity assay of human glioma by scintillation autofluorography. In: Paoletti P, Walker MD, Butti G and Knerich R (eds) Multidisciplinary aspects of brain tumor therapy, Elsevier/Biomedical Press, North-Holland, 1979, pp 19–34
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This paper is part of G. Nikkhah's thesis.
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Nikkhah, G., Tonn, J.C., Hoffmann, O. et al. The MTT assay for chemosensitivity testing of human tumors of the central nervous system. J Neuro-Oncol 13, 1–11 (1992). https://doi.org/10.1007/BF00172941
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DOI: https://doi.org/10.1007/BF00172941