Abstract
We tested the ability of hormones and growth factors to enhance the colony formation in soft agarose of breast carcinoma using two human breast carcinoma cell lines, MCF-7 and MDA-MB231, MCF-7 could clone in a basal medium supplemented only by insulin, transferrin, prostaglandin F2 alpha, and fibronectin. Combining oestradiol, dexamethasone, insulin, transferrin, and triiodothyronine with a basal medium supplemented with 5% (v/v) foetal bovine serum (FBS) increased colony forming efficiency (CFE) two-to three-fold over the best obtained in serum supplemented medium without hormones. While optimal CFE was seen in the hormonally supplemented medium plus 5% FBS, clonal anchorage independent growth could also be obtained without serum for both cell lines by substituting 0.5-1% (v/v) bovine serum albumin (BSA) for FBS. Although CFE was enhanced with the addition of hormones, they did not substantially alter the in vitro chemosensitivity patterns of the cell lines to 8 cytotoxic drugs. Hormonally-supplemented medium with 5% FBS increased the CFE of a small number of fresh specimens of human breast cancer compared with medium supplemented with serum alone. The systematic study of requirements for the in vitro growth of human breast cancer may improve drug sensitivity testing by increasing our ability to grow this neoplasm in culture.
Full text
PDF





Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- 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]
- Alberts D. S., Chen H. S. Tabular summary of pharmacokinetic parameters relevant to in vitro drug assay. Prog Clin Biol Res. 1980;48:351–359. [PubMed] [Google Scholar]
- Allegra J. C., Lippman M. E. Growth of a human breast cancer cell line in serum-free hormone-supplemented medium. Cancer Res. 1978 Nov;38(11 Pt 1):3823–3829. [PubMed] [Google Scholar]
- Ayres K. N. High cloning efficiency of human lymphoid cells in agarose without feeder layers. J Natl Cancer Inst. 1982 Jun;68(6):919–923. [PubMed] [Google Scholar]
- Barnes D., Sato G. Growth of a human mammary tumour cell line in a serum-free medium. Nature. 1979 Oct 4;281(5730):388–389. doi: 10.1038/281388a0. [DOI] [PubMed] [Google Scholar]
- Barnes D., Sato G. Methods for growth of cultured cells in serum-free medium. Anal Biochem. 1980 Mar 1;102(2):255–270. doi: 10.1016/0003-2697(80)90151-7. [DOI] [PubMed] [Google Scholar]
- Barnes D., van der Bosch J., Masui H., Miyazaki K., Sato G. The culture of human tumor cells in serum-free medium. Methods Enzymol. 1981;79(Pt B):368–391. doi: 10.1016/s0076-6879(81)79050-5. [DOI] [PubMed] [Google Scholar]
- 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]
- Cailleau R., Young R., Olivé M., Reeves W. J., Jr Breast tumor cell lines from pleural effusions. J Natl Cancer Inst. 1974 Sep;53(3):661–674. doi: 10.1093/jnci/53.3.661. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Carney D. N., Bunn P. A., Jr, Gazdar A. F., Pagan J. A., Minna J. D. Selective growth in serum-free hormone-supplemented medium of tumor cells obtained by biopsy from patients with small cell carcinoma of the lung. Proc Natl Acad Sci U S A. 1981 May;78(5):3185–3189. doi: 10.1073/pnas.78.5.3185. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Carney D. N., Gazdar A. F., Bunn P. A., Jr, Guccion J. G. Demonstration of the stem cell nature of clonogenic tumor cells from lung cancer patients. Stem Cells. 1982;1(3):149–164. [PubMed] [Google Scholar]
- 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]
- Carney D. N., Mitchell J. B., Kinsella T. J. In vitro radiation and chemotherapy sensitivity of established cell lines of human small cell lung cancer and its large cell morphological variants. Cancer Res. 1983 Jun;43(6):2806–2811. [PubMed] [Google Scholar]
- 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]
- Darfler F. J., Insel P. A. Clonal growth of lymphoid cells in serum-free media requires elimination of H2O2 toxicity. J Cell Physiol. 1983 Apr;115(1):31–36. doi: 10.1002/jcp.1041150106. [DOI] [PubMed] [Google Scholar]
- Elson D. L., Osborne C. K., Livingston R. B., Von Hoff D. D. Methods for determining chemosensitivity of MCF-7 cells in double-layer agar culture: labeling index depression and colony count inhibition. Stem Cells. 1982;2(1):34–44. [PubMed] [Google Scholar]
- Horwitz K. B., Zava D. T., Thilagar A. K., Jensen E. M., McGuire W. L. Steroid receptor analyses of nine human breast cancer cell lines. Cancer Res. 1978 Aug;38(8):2434–2437. [PubMed] [Google Scholar]
- Kaplan P. L., Anderson M., Ozanne B. Transforming growth factor(s) production enables cells to grow in the absence of serum: an autocrine system. Proc Natl Acad Sci U S A. 1982 Jan;79(2):485–489. doi: 10.1073/pnas.79.2.485. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Katz R. L., Lukeman J. M. The comparative diagnostic accuracy of cancer-cell detection obtained with Ficoll-Hypaque gradient separation and standard centrifugation technics on body-cavity fluids. Am J Clin Pathol. 1980 Jul;74(1):18–24. doi: 10.1093/ajcp/74.1.18. [DOI] [PubMed] [Google Scholar]
- Klevjer-Anderson P., Buehring G. C. Effect of hormones on growth rates of malignant and nonmalignant human mammary epithelia in cell culture. In Vitro. 1980 Jun;16(6):491–501. doi: 10.1007/BF02626462. [DOI] [PubMed] [Google Scholar]
- Lewin B. Alternatives for splicing: an intron-coded protein. Cell. 1980 Dec;22(3):645–646. doi: 10.1016/0092-8674(80)90538-3. [DOI] [PubMed] [Google Scholar]
- 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]
- McClure D. B. Anchorage-independent colony formation of SV40 transformed BALB/c-3T3 cells in serum-free medium: role of cell- and serum-derived factors. Cell. 1983 Mar;32(3):999–1006. doi: 10.1016/0092-8674(83)90084-3. [DOI] [PubMed] [Google Scholar]
- Pathak M. A., Matrisian L. M., Magun B. E., Salmon S. E. Effect of epidermal growth factor on clonogenic growth of primary human tumor cells. Int J Cancer. 1982 Dec 15;30(6):745–750. doi: 10.1002/ijc.2910300611. [DOI] [PubMed] [Google Scholar]
- 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]
- 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]
- 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]
- Sandbach J., Von Hoff D. D., Clark G., Cruz A. B., Jr, Obrien M. Direct cloning of human breast cancer in soft agar culture. Cancer. 1982 Oct 1;50(7):1315–1321. doi: 10.1002/1097-0142(19821001)50:7<1315::aid-cncr2820500717>3.0.co;2-7. [DOI] [PubMed] [Google Scholar]
- 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]
- Simms E., Gazdar A. F., Abrams P. G., Minna J. D. Growth of human small cell (oat cell) carcinoma of the lung in serum-free growth factor-supplemented medium. Cancer Res. 1980 Dec;40(12):4356–4363. [PubMed] [Google Scholar]
- 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]
- Soule H. D., Vazguez J., Long A., Albert S., Brennan M. A human cell line from a pleural effusion derived from a breast carcinoma. J Natl Cancer Inst. 1973 Nov;51(5):1409–1416. doi: 10.1093/jnci/51.5.1409. [DOI] [PubMed] [Google Scholar]
- Tveit K. M., Endresen L., Rugstad H. E., Fodstad O., Pihl A. Comparison of two soft-agar methods for assaying chemosensitivity of human tumours in vitro: malignant melanomas. Br J Cancer. 1981 Oct;44(4):539–544. doi: 10.1038/bjc.1981.223. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Von Hoff D. D., Casper J., Bradley E., Sandbach J., Jones D., Makuch R. Association between human tumor colony-forming assay results and response of an individual patient's tumor to chemotherapy. Am J Med. 1981 May;70(5):1027–1041. doi: 10.1016/0002-9343(81)90859-7. [DOI] [PubMed] [Google Scholar]
