Efficient fixation procedure of human leukemia cells in sulforhodamine B cytotoxicity assay

doi:10.1016/S1056-8719(96)00113-X

Journal of Pharmacological and Toxicological Methods

Volume 36, Issue 3, November 1996, Pages 163-169

https://doi.org/10.1016/S1056-8719(96)00113-X Get rights and content

Abstract

The fixation procedures in sulforhodamine B (SRB) assay for human leukemia cells were modified to produce more reliable results. It was found that the concentration of the fixative agent, trichloroacetic acid (TCA), was critical in the selective fixation of cellular protein. While a TCA solution of 80% fixed both cells and serum proteins, a 50% solution fixed only cells with a very low interference of the serum proteins. Accordingly, we selected 50% TCA as a fixative agent which made the final absorbance of the SRB assay to be exactly matched to the cell density with a small deviation and a low background. Besides the change of TCA concentration, a precentifugation of microplate just before fixation also improved the previous assay procedures in the two points of view. The 2-h standing step was simply substituted for only 1 min of centrifugation. Both the rapid and slow application of TCA solution in fixation produced the same extents of fixation. In an actual application, these two kinds of modifications in the previous SRB assay procedure were also proved to be effective in the determination of cytotoxicities of doxorubicin by using human leukemias.

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^☆: This work was supported by HAN project of Ministry of Science and Technology, Korea.

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Original articleEfficient fixation procedure of human leukemia cells in sulforhodamine B cytotoxicity assay☆

Abstract

Biochem Pharmacol

J Immunol Methods

Feasibility of drug screening with panels of human tumor cell lines using a microculture tetrazolium assay

Cancer Res

Status of the National Cancer Institute preclinical antitumor drug discovery screen

Growth and chemotherapeutic response of cells in a hollow-fiber in vitro solid tumor model

J Natl Cancer Inst

Antiproliferative effect of nonsteroidal antiinflammatory drugs against human colon cancer cells

Cancer Epidemiol Biomarkers Prev

Establishment and characterization of an in vitro model of acquired resistance to cisplatin in a human testicular nonseminomatous germ cell line

Cancer Res

Original article
Efficient fixation procedure of human leukemia cells in sulforhodamine B cytotoxicity assay☆