Measurement of human tumour cell growth in soft-agar cultures using computer-assisted volume analysis

Alley, Michael C.; Lieber, Michael M.

doi:10.1038/bjc.1985.179

Br J Cancer

1985

DOI: 10.1038/bjc.1985.179

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Measurement of human tumour cell growth in soft-agar cultures using computer-assisted volume analysis

Michael C. Alley¹,

Michael M. Lieber²

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1985

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Cited by 6 publications

(1 citation statement)

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“…This extensive experience has generated concern about technical problems which occur in the performance of soft agar colony forming assays using samples of cells isolated from primary human tumours; publications on this topic have appeared in this journal previously (Agrez et al, 1982a; Alley & Lieber 1984& Lieber , 1985. Consequently, it was of interest to investigate whether the use of a different way of assessing cell proliferation and drug effects in such soft agar tumour colony forming assays might be as effective or more effective than simply optically counting the number of colonies formed.…”

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Soft agarose culture human tumour colony forming assay for drug sensitivity testing: [3H]-thymidine incorporation vs colony counting

Jones¹,

Tsukamoto²,

O’Brien³

et al. 1985

Br J Cancer

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confidence: 99%

Soft agarose culture human tumour colony forming assay for drug sensitivity testing: [3H]-thymidine incorporation vs colony counting

Jones¹,

Tsukamoto²,

O’Brien³

et al. 1985

Br J Cancer

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The determination of growth rates of individual colonies in agarose using high‐resolution automated image analysisx

et al. 1990

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This paper describes the evaluation of a colony formation assay using automated image analysis, which permits the tracking of growth at the individual colony level, such that a growth rate can be estimated for each colony followed. In principle, this will permit quantitative characterization of cellular heterogeneity in growth rate and cellular heterogeneity in response to proliferation-modifying agents. In addition, we have demonstrated the possibility of using correlative microscopy to relate growth rate to other parameters, using metabolic viability as an example. This should be useful for determining cellular characteristics associated with proliferative behavior and response to proliferationmodifying agents.Key terms: Proliferation rate, correlative microscopy, tetrazolium dye, growth factors, antiproliferative drugs, drug response, computer simulation, heterogeneityThe presence of variant cells within a population derived from a common ancestor is known as cellular heterogeneity (8,27). This situation represents a significant problem in cancer therapeutics, for example, in that diversely responsive subpopulations within a target tissue may allow for selection of resistant cells during therapy (5). To study cellular heterogeneity directly, it is necessary to have methods that are capable of defining cellular characteristics and behavior a t the level ofthe individual cell. Advances in flow cytometry and image analysis are making such studies increasingly possible.Proliferation is an important cell behavior in cancer research, inasmuch as cancer is primarily a disease of growth control, and anticancer therapies are usually aimed at killing, or otherwise arresting the growth of, malignant cells. Unusual cell proliferation also plays a role in arteriosclerosis (7), and thus is a cell behavior with relevance to more than one significant health problem. Although cytometric systems, such as flow approaches, permit multiple parameter measurements on individual cells virtually instantaneously, such measurements do not permit quantification of cell proliferation.A method commonly used for evaluation of proliferative potential of individual cells is the colony formation assay, wherein immobilized cells are allowed to proliferate, and growth is quantified by enumeration of progeny. Computer-assisted evaluation of colony formation assays has been employed for colony counting (10,141, for total colony volume analysis (29,311, and for histogram analysis of colony sizes (12). These approaches permit assessment of growth kinetics of populations and estimation of growth rate of individual colonies by inference. A manual approach involving colony isolation and cell enumeration has been used to determine numbers of cells in colonies in soft agar (29); a similar approach has been used to describe heterogeneity in growth rate among individual colonies growing on plastic (15). Time-lapse cinematographic approaches (1 1,211 and time-lapse video approaches (6,7) have been used to determine the growth rate of individual cells throu...

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In vitro assays for new drug screening: Comparison of a thymidine incorporation assay with the human tumor colony‐forming assay

Hildebrand-Zanki

1987

The International Journal of Cell Cloning

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Because of technical limitations with the human tumor colony-forming assay (HTCFA), we determined the feasibility of using a thymidine incorporation assay (TIA) for new drug screening. Concordance between the TIA and the HTCFA was obtained (r = 0.840) with twenty coded compounds. Toxic agents without proven clinical efficacy were active in both assays, while non-toxic substances were inactive. Growth rates were higher with the TIA (75%) than with the HTCFA (45%), and the TIA could be completed in 6 days compared to 14-21 days with the HTCFA. We concluded that the TIA is a useful adjunct to in vivo tumor models for screening new anticancer agents.

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Measurement of human tumour cell growth in soft-agar cultures using computer-assisted volume analysis

Cited by 6 publications

References 8 publications

Soft agarose culture human tumour colony forming assay for drug sensitivity testing: [3H]-thymidine incorporation vs colony counting

Soft agarose culture human tumour colony forming assay for drug sensitivity testing: [3H]-thymidine incorporation vs colony counting

The determination of growth rates of individual colonies in agarose using high‐resolution automated image analysisx

In vitro assays for new drug screening: Comparison of a thymidine incorporation assay with the human tumor colony‐forming assay

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