This Article Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Alley, M. C. Articles by Boyd, M. R. Search for Related Content PubMed PubMed Citation Articles by Alley, M. C. Articles by Boyd, M. R.

Morphometric and Colorimetric Analyses of Human Tumor Cell Line Growth and Drug Sensitivity in Soft Agar Culture¹

Laboratory of Drug Discovery Research and Development, Developmental Therapeutics Program [M. C. A., C. M. P., M. L. H., M. R. B.], and Biometric Research Branch, Cancer Therapy Evaluation Program [L. R. R.], Division of Cancer Treatment, National Cancer Institute, Frederick Cancer Research and Development Center, Frederick, Maryland 21701-1013

Previous studies have demonstrated the suitability of image analysis of tetrazolium-stained colonies to assess growth and drug sensitivity of human tumor cells cultivated in soft agar culture. In the present study, the potential utility of colorimetric analysis to expedite experimental drug evaluations using human tumor cell lines was investigated. The same culture dishes were assessed by image analysis and by formazan colorimetry for purposes of comparing multiple methods of measuring growth as well as growth inhibition. Replicate cultures treated with 2-(p-iodonitrophenyl)-3-p-nitrophenyl-5-phenyltetrazolium chloride or 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide exhibited nearly identical colony count and volume indices as well as excellent correlation in colorimetric end points. Colony-forming unit volume analysis versus colorimetric assessment of the same cultures following dimethyl sulfoxide extraction of protamine sulfate-rinsed, dried soft agar cultures exhibited excellent linear correlation for both growth (Pearson r ranging from 0.95 to 1.00) and drug sensitivity (Pearson r ranging from 0.90 to 0.99, and Spearman r ranging from 0.82 to 0.97) and similar drug sensitivity profiles. Results of the current investigation indicate that end points of soft agar culture remain stable for a period of at least 2 weeks following assay termination. In addition, a colorimetric detection range of 1.3–2.2 log units permits determinations of survival levels ranging from 100 to 5% of respective control levels. Colorimetric analysis is anticipated to expedite soft agar colony formation assay evaluations (a) by reducing the need to use the more rigorous and time-consuming image analysis procedures to measure activity in preliminary drug sensitivity assays and (b) by permitting the determination of effective concentration ranges of new experimental agents for subsequent, more detailed investigations.

¹ Research supported in part with Federal funds from the Department of Health and Human Services under Contract NO1-CO-74102. The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the United States Government.

² To whom requests for reprints should be addressed.

Received 5/ 8/90. Accepted 12/ 4/90.

This article has been cited by other articles:

				M. G. Hollingshead, J. P. Carter, K. Dougherty, and C. A. Bonomi The Hollow Fiber Assay in Cancer Efficacy Testing in the Developmental Therapeutics Program at the National Cancer Institute Am. Assoc. Cancer Res. Educ. Book, April 1, 2005; 2005(1): 351 - 354. [Full Text] [PDF]

				J. A. Hartley, V. J. Spanswick, N. Brooks, P. H. Clingen, P. J. McHugh, D. Hochhauser, R. B. Pedley, L. R. Kelland, M. C. Alley, R. Schultz, et al. SJG-136 (NSC 694501), a Novel Rationally Designed DNA Minor Groove Interstrand Cross-Linking Agent with Potent and Broad Spectrum Antitumor Activity: Part 1: Cellular Pharmacology, In vitro and Initial In vivo Antitumor Activity Cancer Res., September 15, 2004; 64(18): 6693 - 6699. [Abstract] [Full Text] [PDF]

				G. P. Dasmahapatra, P. Didolkar, M. C. Alley, S. Ghosh, E. A. Sausville, and K. K. Roy In vitro Combination Treatment with Perifosine and UCN-01 Demonstrates Synergism against Prostate (PC-3) and Lung (A549) Epithelial Adenocarcinoma Cell Lines Clin. Cancer Res., August 1, 2004; 10(15): 5242 - 5252. [Abstract] [Full Text] [PDF]

				M. D. Sadar, V. A. Akopian, and E. Beraldi Characterization of a New in Vivo Hollow Fiber Model for the Study of Progression of Prostate Cancer to Androgen Independence Mol. Cancer Ther., June 1, 2002; 1(8): 629 - 637. [Abstract] [Full Text] [PDF]

				E. A. SAUSVILLE, J. JOHNSON, M. ALLEY, D. ZAHAREVITZ, and A. M. SENDEROWICZ Inhibition of CDKs as a Therapeutic Modality Ann. N.Y. Acad. Sci., June 1, 2000; 910(1): 207 - 222. [Abstract] [Full Text] [PDF]

				M. Kobayashi, S. Nagata, T. Iwasaki, K. Yanagihara, I. Saitoh, Y. Karouji, S. Ihara, and Y. Fukui Dedifferentiation of adenocarcinomas by activation of phosphatidylinositol 3-kinase PNAS, April 27, 1999; 96(9): 4874 - 4879. [Abstract] [Full Text] [PDF]

				T. Akagi, T. Shishido, K. Murata, and H. Hanafusa v-Crk activates the phosphoinositide 3-kinase/AKT pathway in transformation PNAS, June 20, 2000; 97(13): 7290 - 7295. [Abstract] [Full Text] [PDF]