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British Journal of Cancer logoLink to British Journal of Cancer
. 1984 Feb;49(2):225–233. doi: 10.1038/bjc.1984.35

Improved optical detection of colony enlargement and drug cytotoxicity in primary soft agar cultures of human solid tumour cells.

M C Alley, M M Lieber
PMCID: PMC1976703  PMID: 6199034

Abstract

The presence of cellular aggregates in cell suspensions derived from human solid tumours often complicates subsequent evaluation of colony formation in primary soft agar cultures (Agrez et al., 1982b). In the present study, performance of a conventional colony formation assay was observed to lack sufficient sensitivity to identify growth and active chemotherapeutic agents in the majority of specimen cultures. Modification of conventional methodologies to include filtration of cell suspensions, use of "proliferation control" and "cytotoxicity control" cultures as well as vital staining were found to be essential for the valid assessment of primary soft agar cultures in our laboratory. In addition, application of drugs to culture surface in place of culture incorporation appeared to facilitate culture performance and drug sensitivity testing.

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Selected References

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  1. Agrez M. V., Kovach J. S., Beart R. W., Jr, Rubin J., Moertel C. G., Lieber M. M. Human colorectal carcinoma: patterns of sensitivity to chemotherapeutic agents in the human tumor stem cell assay. J Surg Oncol. 1982 Jul;20(3):187–191. doi: 10.1002/jso.2930200311. [DOI] [PubMed] [Google Scholar]
  2. Agrez M. V., Kovach J. S., Lieber M. M. Cell aggregates in the soft agar "human tumour stem-cell assay". Br J Cancer. 1982 Dec;46(6):880–887. doi: 10.1038/bjc.1982.297. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Alley M. C., Uhl C. B., Lieber M. M. Improved detection of drug cytotoxicity in the soft agar colony formation assay through use of a metabolizable tetrazolium salt. Life Sci. 1982 Dec 27;31(26):3071–3078. doi: 10.1016/0024-3205(82)90077-7. [DOI] [PubMed] [Google Scholar]
  4. Bertoncello I., Bradley T. R., Campbell J. J., Day A. J., McDonald I. A., McLeish G. R., Quinn M. A., Rome R., Hodgson G. S. Limitations of the clonal agar assay for the assessment of primary human ovarian tumour biopsies. Br J Cancer. 1982 Jun;45(6):803–811. doi: 10.1038/bjc.1982.131. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bol S., van den Engh G., Visser J. A technique for staining haemopoietic colonies in agar cultures. Exp Hematol. 1977 Nov;5(6):551–553. [PubMed] [Google Scholar]
  6. Hamburger A. W., Salmon S. E. Primary bioassay of human tumor stem cells. Science. 1977 Jul 29;197(4302):461–463. doi: 10.1126/science.560061. [DOI] [PubMed] [Google Scholar]
  7. Hamburger A., Salmon S. E. Primary bioassay of human myeloma stem cells. J Clin Invest. 1977 Oct;60(4):846–854. doi: 10.1172/JCI108839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Jerne N. K., Henry C., Nordin A. A., Fuji H., Koros A. M., Lefkovits I. Plaque forming cells: methodology and theory. Transplant Rev. 1974;18:130–191. doi: 10.1111/j.1600-065x.1974.tb01588.x. [DOI] [PubMed] [Google Scholar]
  9. Lieber M. M., Kovach J. S. Soft agar colony formation assay for chemotherapy sensitivity testing of human solid tumors. Mayo Clin Proc. 1982 Aug;57(8):527–528. [PubMed] [Google Scholar]
  10. Rupniak H. T., Hill B. T. The poor cloning ability in agar of human tumour cells from biopsies of primary tumours. Cell Biol Int Rep. 1980 May;4(5):479–486. doi: 10.1016/0309-1651(80)90035-1. [DOI] [PubMed] [Google Scholar]
  11. Schaeffer W. I., Friend K. Efficient detection of soft agar grown colonies using a tetrazolium salt. Cancer Lett. 1976 May;1(5):259–262. doi: 10.1016/s0304-3835(75)97506-0. [DOI] [PubMed] [Google Scholar]
  12. Selby P., Buick R. N., Tannock I. A critical appraisal of the "human tumor stem-cell assay". N Engl J Med. 1983 Jan 20;308(3):129–134. doi: 10.1056/NEJM198301203080304. [DOI] [PubMed] [Google Scholar]
  13. Soehnlen B., Young L., Liu R. Standard laboratory procedures for in vitro assay of human tumor stem cells. Prog Clin Biol Res. 1980;48:331–338. [PubMed] [Google Scholar]

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