Test for chemotherapeutic sensitivity of cerebral gliomas: use of colorimetric MTT assay

Abstract

This study was undertaken to evaluate the colorimetric MTT [3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2H tetrazolium bromide] assay as a means of testing the sensitivity of gliomas to chemotherapeutic agents in vitro. Eight human glioma established cell lines were plated in 96-well tissue culture plates and incubated for 4 days with 10 different anti-cancer agents; 5 different concentrations of each drug were tested. The MTT dye was then added to the wells, and the resulting formazan precipitate was solubilized with dimethylsulfoxide (DMSO). The spectrophotometric absorbance (measured at 570 nm) of control and experimental wells was used to calculate the cytotoxicity index (CI). Values with a CI greater than 50% growth inhibition indicated cytotoxic efficacy (sensitivity to the chemotherapeutic drug). Six of the seven (85.7%) glioma cell lines were highly sensitive at varying concentrations to mitomycin C, cisplatin, and doxorubicin. Four of the seven (57.1%) cell lines demonstrated intermediate sensitivity to mitoxantrone and vinblastine. Five of the seven (71.4%) cell lines exhibited resistance to etoposide, bleomycin, cosmegen, and BCNU. One of the cell lines tested, U-138MG, failed to produce the MTT formazan precipitate, so that the sensitivity of this cell line to the panel chemotherapeutic drugs could not be determined. The variability of the results indicates the need for an in vitro screening method to evaluate the effectiveness of clinical and experimental chemotherapeutic agents. The MTT assay provides a rapid method of screening antineoplastic agents against gliomas for cytotoxicity.

Flow Cytometry Analysis of Neural Differentiation Markers Expression in Human Glioblastomas May Predict Their Response to Chemotherapy

“…However, each assay has pitfalls such as artificial selective pressure inherent to tissue culture sytem, heterogeneity of biopsy specimen, in vitro drifting of cultured cells from homogeneous to heteroclonal culture, possible cytotoxic effect of metabolites, not the parent compound, and finally heterogeneity in chemosensitivity and consecutive problems with the interpretation of results (Jordan et al 1992;Kimmel et al 1987;Yung 1989). While expression of the above-mentioned markers has been correlated with tumour malignancy, proliferative activity, invasive potential or patient prognosis, there is only a limited knowledge about such associations with respect to the sensitivity of astrocytic tumours to chemotherapy.…”

Chemosensitivity of glioma cells in vitro: a meta analysis

“…The most frequently used test was the MTT test (Hansen et al 1989;Kuratsu et al 1991;Jordan et al 1992;Wol and JuÈ rgens 1994;Wol et al 1996;Poppenborg et al 1997), the most sensitive tests were the colonyforming assays ( Table 1).…”

O-Benzyl-N-(9-acridinyl)hydroxylamines

“…The reduction of MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-2H tetrazolium bromide), a yellow tetrazole, to its insoluble formazan was observed and resulted in a color change that was easily monitored by a change in absorbance at 560 nm. 16 The reduction of the dye is dependent on NAD(P)H-dependent oxidoreductase enzymes that are found in large amounts within the cytosolic compartment of the cell. These enzymes are indicative of the number of viable cells within a sample as they are tied to cell proliferation.…”