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[Cancer Research 47, 936-942, February 15, 1987]
© 1987 American Association for Cancer Research
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Evaluation of a Tetrazolium-based Semiautomated Colorimetric Assay: Assessment of Chemosensitivity Testing1

James Carmichael2, William G. DeGraff, Adi F. Gazdar, John D. Minna and James B. Mitchell

NCI-Navy Medical Oncology Branch [J.C., A.F.G., J.D.M.] and Radiobiology Section, Radiation Oncology Branch [J.C., W.G.D., J.B.M.], National Cancer Institute and Naval Hospital, Bethesda, Maryland 20814

Drug sensitivity assays were performed using a variation of a colorimetric [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)] assay on V79, CHO-AuxB1, CHRC5, NCI-H460, and NCI-H249 cell lines following optimization of experimental conditions for each cell line. Results from this assay were compared with data assimilated simultaneously by clonogenic assay and by dye exclusion assay. Good correlation was observed using the CHO-AuxB1 cell line and the pleiotropic drug-resistant mutant CHRC5, with similar degrees of relative resistance observed with both the MTT and clonogenic assays. Good correlation was observed between the clonogenic and MTT assays for 1-h drug exposures, although the MTT assay was more sensitive to vinblastine. In general, the clonogenic assay was more sensitive when continuous drug exposures were utilized, although this was primarily related to the increased drug exposure time. While the use of the MTT assay in drug sensitivity testing of primary tumor samples is limited, since contaminating normal cells may also reduce the tetrazolium, the MTT assay can be semiautomated, and therefore it offers a valid, simple method of assessing chemosensitivity in established cell lines.

1 The opinions or assertions contained herein are the private views of the authors and are not to be construed as official or as reflecting the views of the Department of the Navy or the Department of Defense.

2 To whom requests for reprints should be addressed, at Radiation Oncology Branch, National Cancer Institute, Bldg. 10, Room B3-B69, Bethesda, MD 20892.

Received 4/25/86. Revised 9/18/86. Accepted 10/30/86.




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