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. 1982 Jun;45(6):803–811. doi: 10.1038/bjc.1982.131

Limitations of the clonal agar assay for the assessment of primary human ovarian tumour biopsies.

I Bertoncello, T R Bradley, J J Campbell, A J Day, I A McDonald, G R McLeish, M A Quinn, R Rome, G S Hodgson
PMCID: PMC2011038  PMID: 7093117

Abstract

114 biopsy specimens from 70 patients with ovarian carcinoma at all stages of disease were submitted for assessment of clonogenic capacity in agar. A highly significant correlation was found between agar clonogenicity and patient survival after biopsy. However, problems related to inherent tumour heterogeneity, quality of sample and tissue disaggregation indicate that this technique may have limited applicability in the routine assessment of patients. Only 41 biopsy specimens (36%) from 31 patients (44.3%) complied with the prerequisite criteria for agar clonogenic assessment, namely: (a) the confirmed presence of malignant cells in the biopsy, (b) the ability to prepare a single-cell suspension, and (c) adequate viable cell numbers for assay. Furthermore, although the dominant patterns of agar clonogenic growth could be identified and correlated with stage of disease, the heterogeneity in both initial clonogenic capacity and "self-renewal" capacity assessed by the ability of primary clones to propagate in liquid culture and reclone in agar was too inconsistent for the assay to be used as a prognostic index for the individual patient.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Alberts D. S., Samon S. E., Chen H. S., Surwit E. A., Soehnlen B., Young L., Moon T. E. In-vitro clonogenic assay for predicting response of ovarian cancer to chemotherapy. Lancet. 1980 Aug 16;2(8190):340–342. doi: 10.1016/s0140-6736(80)90340-2. [DOI] [PubMed] [Google Scholar]
  2. Barrett J. C., Ts'o P. O. Evidence for the progressive nature of neoplastic transformation in vitro. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3761–3765. doi: 10.1073/pnas.75.8.3761. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bradley T. R., Hodgson G. S., Rosendaal M. The effect of oxygen tension on haemopoietic and fibroblast cell proliferation in vitro. J Cell Physiol. 1978 Dec;97(3 Pt 2 Suppl 1):517–522. doi: 10.1002/jcp.1040970327. [DOI] [PubMed] [Google Scholar]
  4. Buick R. N., Fry S. E., Salmon S. E. Effect of host-cell interactions on clonogenic carcinoma cells in human malignant effusions. Br J Cancer. 1980 May;41(5):695–704. doi: 10.1038/bjc.1980.131. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Carney D. N., Gazdar A. F., Minna J. D. Positive correlation between histological tumor involvement and generation of tumor cell colonies in agarose in specimens taken directly from patients with small-cell carcinoma of the lung. Cancer Res. 1980 Jun;40(6):1820–1823. [PubMed] [Google Scholar]
  6. Courtenay V. D., Mills J. An in vitro colony assay for human tumours grown in immune-suppressed mice and treated in vivo with cytotoxic agents. Br J Cancer. 1978 Feb;37(2):261–268. doi: 10.1038/bjc.1978.35. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Courtenay V. D., Selby P. J., Smith I. E., Mills J., Peckham M. J. Growth of human tumour cell colonies from biopsies using two soft-agar techniques. Br J Cancer. 1978 Jul;38(1):77–81. doi: 10.1038/bjc.1978.165. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Hamburger A. W., Salmon S. E., Kim M. B., Trent J. M., Soehnlen B. J., Alberts D. S., Schmidt H. J. Direct cloning of human ovarian carcinoma cells in agar. Cancer Res. 1978 Oct;38(10):3438–3444. [PubMed] [Google Scholar]
  9. 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]
  10. Hard G. C., King H., Borland R., Stewart B. W. Length of in vivo exposure to a carcinogenic dose of dimethylnitrosamine necessary for subsequent expression of morphological transformation by rat kidney cells in vitro. Oncology. 1977;34(1):16–19. doi: 10.1159/000225174. [DOI] [PubMed] [Google Scholar]
  11. Ioachim H. L., Sabbath M., Andersson B., Barber H. R. Tissue cultures of ovarian carcinomas. Lab Invest. 1974 Oct;31(4):381–390. [PubMed] [Google Scholar]
  12. Laboisse C. L., Augeron C., Potet F. Growth and differentiation of human gastrointestinal adenocarcinoma stem cells in soft agarose. Cancer Res. 1981 Jan;41(1):310–315. [PubMed] [Google Scholar]
  13. Mouriquand J., Mouriquand C., Petitpas E., Mermet M. A. Long-term tissue cultures of human pleural effusions: a cytological follow-up. In Vitro. 1978 Jul;14(7):591–600. doi: 10.1007/BF02617918. [DOI] [PubMed] [Google Scholar]
  14. Neugut A. I., Weinstein I. B. The use of agarose in the determination of anchorage-independent growth. In Vitro. 1979 May;15(5):351–355. doi: 10.1007/BF02616141. [DOI] [PubMed] [Google Scholar]
  15. Nowell P. C. The clonal evolution of tumor cell populations. Science. 1976 Oct 1;194(4260):23–28. doi: 10.1126/science.959840. [DOI] [PubMed] [Google Scholar]
  16. PUCK T. T., MARCUS P. I. Action of x-rays on mammalian cells. J Exp Med. 1956 May 1;103(5):653–666. doi: 10.1084/jem.103.5.653. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Pavelic Z. P., Slocum H. K., Rustum Y. M., Creaven P. J., Nowak N. J., Karakousis C., Takita H., Mittelman A. Growth of cell colonies in soft agar from biopsies of different human solid tumors. Cancer Res. 1980 Nov;40(11):4151–4158. [PubMed] [Google Scholar]
  18. Roscoe J. P., Hince T. A., Claisse P. J., Winslow D. P. Effect of 12-O-tetradecanoylphorbol-13-acetate on two charateristics of transformation acquired sequentially by ENU-exposed rat brain cells. Br J Cancer. 1980 Nov;42(5):756–764. doi: 10.1038/bjc.1980.309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. 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]
  20. Salmon S. E., Hamburger A. W., Soehnlen B., Durie B. G., Alberts D. S., Moon T. E. Quantitation of differential sensitivity of human-tumor stem cells to anticancer drugs. N Engl J Med. 1978 Jun 15;298(24):1321–1327. doi: 10.1056/NEJM197806152982401. [DOI] [PubMed] [Google Scholar]
  21. Selby P. J., Raghavan D. Role of laboratory chemosensitivity testing in the selection of cancer chemotherapy for individual patients. J Clin Pathol. 1981 May;34(5):455–463. doi: 10.1136/jcp.34.5.455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Shin S. I., Freedman V. H., Risser R., Pollack R. Tumorigenicity of virus-transformed cells in nude mice is correlated specifically with anchorage independent growth in vitro. Proc Natl Acad Sci U S A. 1975 Nov;72(11):4435–4439. doi: 10.1073/pnas.72.11.4435. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. TOLMACH L. J., MARCUS P. I. Development of x-ray induced giant HeLa cells. Exp Cell Res. 1960 Aug;20:350–360. doi: 10.1016/0014-4827(60)90163-4. [DOI] [PubMed] [Google Scholar]
  24. Von Hoff D. D., Casper J., Bradley E., Trent J. M., Hodach A., Reichert C., Makuch R., Altman A. Direct cloning of human neuroblastoma cells in soft agar culture. Cancer Res. 1980 Oct;40(10):3591–3597. [PubMed] [Google Scholar]

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