Skip to main content
SpringerLink
Log in

Selective growth of freshly isolated human breast epithelial cells cultured at low concentrations in the presence or absence of bone marrow cells

  • Report
  • Published:
Breast Cancer Research and Treatment Aims and scope Submit manuscript

Summary

In this study, we show that conditions previously found to promote the selective growth of human breast epithelial cells (HBEC) in serum-free primary cultures established from normal or malignant tissue can be extended to cultures initiated at low seeding densities (< 5000 cells/cm2). The epithelial nature of the cells produced was documented by their positive staining with antibodies specific for keratins 8, 14, and 18, and 2 antibodies that recognize epithelial-specific antigens (Ber-EP4 and HB8630). HBEC growth was not affected, either positively or negatively, by the use of a medium containing a combination of fetal calf and horse serum, which promotes the growth of many types of stromal cells and associated hematopoietic precursors, or by the inclusion in the initial cell suspension of marrow cells at HBEC to marrow cell ratios typical of bone marrow samples from patients with metastatic breast cancer. The presence of fibroblast feeders from a variety of sources enhanced the growth of HBEC to different degrees. In cultures initiated with low numbers of cells obtained from samples of breast carcinoma, HBEC growth was generally reduced by comparison to cultures of normal HBEC. With the detection methods used, it was not possible to determine the extent to which this decreased growth was due to a reduced frequency of malignant HBEC within vitro precursor activity, or the presence of reduced numbers of residual normal HBEC precursors, or both. However, preliminary data indicate that this approach also allows the detection of some breast carcinoma cells with proliferative ability that are present in the marrow or pleural effusions of some breast cancer patients. These studies demonstrate the feasibility of detecting normal and malignant HBEC with growth potential when these are cultured at low density and/or as rare contaminants of marrow cell suspensions, and provide a starting point for their further characterization.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

References

  1. Bonadonna G: Conceptual and practical advances in the management of breast cancer. J Clin Oncol 7: 1380–1397, 1989

    PubMed  Google Scholar 

  2. Harbeck N, Untch M, Pache L, Eiermann W: Tumour cell detection in the bone marrow of breast cancer patients at primary therapy: results of a 3-year median follow-up. Br J Cancer 69: 566–571, 1994

    PubMed  Google Scholar 

  3. Antman KH: Dose-intensive therapy in breast cancer. In: Armitage JO, Antman KH (Eds) High-Dose Cancer Therapy. Pharmacology, Hematopoietins, Stem Cells. Williams & Wilkins, Baltimore, Maryland, 1992, pp 701–718

    Google Scholar 

  4. Sulkes A: The role of high-dose chemotherapy with autologous bone marrow support in breast cancer: state of the art. Isr J Med Sci 29: 111–116, 1993

    PubMed  Google Scholar 

  5. Peters WP, Ross M, Vredenburgh JJ, Meisenberg B, Marks LB, Winer E, Kurtzberg J, Bast RC Jr, Jones J, Shpall E, Wu K, Rosner G, Gilbert C, mathias B, Coniglio D, Petros W, Henderson IC, Norton L, Weiss RB, Budman D, Hurd D: High-dose chemotherapy and autologous bone marrow support as consolidation after standard-dose adjuvant therapy for high-risk primary breast cancer. J Clin Oncol 11: 1132–1143, 1993

    PubMed  Google Scholar 

  6. Kennedy MJ, Davis J, Passos-Coelho J, Noga SJ, Huelskamp AM, Ohly K, Davidson NE: Administration of human recombinant granulocyte colony-stimulating factor (Filgrastim) accelerates granulocyte recovery following high-dose chemotherapy and autologous marrow transplantation with 4-hydroperoxycyclophosphamide-purged marrow in women with metastatic breast cancer. Cancer Res 53: 5424–5428, 1993

    PubMed  Google Scholar 

  7. Brugger W, Bross KJ, Glatt M, Weber F, Mertelsmann R, Kanz L: Mobilization of tumor cells and hematopoietic progenitor cells into peripheral blood of patients with solid tumors. Blood 83: 636–640, 1994

    PubMed  Google Scholar 

  8. Dearnaley DP, Ormerod MG, Soane JP: Micrometastases in breast cancer: long-term follow-up of the first patient cohort. Eur J Cancer 27: 236–239, 1991

    PubMed  Google Scholar 

  9. Sharp JG, Crouse DA: Marrow contamination: detection and significance. In: Armitage JO, Antman KH (eds) High-Dose Cancer Therapy, Pharmacology, Hematopoietins, Stem Cells. Williams and Wilkins, Baltimore, 1992, pp 226–248

    Google Scholar 

  10. Ross AA, Cooper BW, Lazarus HM, Mackay W, Moss TJ, Ciobanu N, Tallman MS, Kenned MJ, Davidson NE, Sweet D, Winter C, Akard L, Jansen J, Copelan E, Meagher RC, Herzig RH, Klumpp TR, Kahn DG, Warner NE: Detection and viability of tumor cells in peripheral blood stem cell collections from breast cancer patients using immunocytochemical and clonogenic assay techniques. Blood 82: 2605–2610, 1993

    PubMed  Google Scholar 

  11. Cote RJ, Rosen PP, Lesser ML, Old LJ, Osborne MP: Prediction of early relapse in patients with operable breast cancer by detection of occult bone marrow micrometastases. J Clin Oncol 9: 1749–1756, 1991

    PubMed  Google Scholar 

  12. Mansi JL, Easton D, Berger U, Gazet JC, Ford HT, Dearnaley D, Coombes RC: Bone marrow micrometastases in primary breast cancer: Prognostic significance after 6 years' follow-up. Eur J Cancer 27: 1552–1555, 1991

    PubMed  Google Scholar 

  13. Courtenay VD, Mills J:In vitro colony assay for human tumours grown in immunosuppressed mice and treatedin vivo with cytotoxic agents. Br J Cancer 37: 261–268, 1978

    PubMed  Google Scholar 

  14. Salmon SE, Hamburger AW, Soehnlen B, Durie BG, Alberts DS, Moon TE: Quantitation of differential sensitivity of human tumor stem cells to anticancer drugs. N Engl J Med 298: 1321–1327, 1978

    PubMed  Google Scholar 

  15. Smith HS, Lan S, Ceriani R, Hackett AJ, Stampfer MR: Clonal proliferation of cultured non-malignant and malignant human breast epithelia. Cancer Res 41: 4637–4643, 1981

    PubMed  Google Scholar 

  16. Cillo C, Odartchenko N: Cloning of human tumor cells in methylcellulose-containing medium. In: Predictive Drug Testing on Human Tumor Cells. Springer, Berlin, Heidelberg, New York, 1984, pp 56–64

    Google Scholar 

  17. Ethier SP, Mahacek ML, Gullick WJ, Frank TS, Weber BL: Differential isolation of normal luminal mammary epithelial cells and breast cancer cells from primary and metastatic sites using selective media. Cancer Res 53: 627–635, 1993

    PubMed  Google Scholar 

  18. Emerman JT, Wilkinson DA: Routine culturing of normal, dysplastic and malignant human mammary epithelial cells from small tissue samples.In Vitro Cell Dev Biol 26: 1186–1194, 1990

    PubMed  Google Scholar 

  19. Lemoine FM, Humphries RK, Abraham SDM, Krystal G, Eaves CJ: Partial characterization of a novel stromal cellderived pre-B-cell growth factor active on normal and immortalized pre-B cells. Exp Hematol 16: 718–726, 1988

    PubMed  Google Scholar 

  20. Eaves CJ, Cashman JD, Eaves AC: Methodology of longterm culture of human hemopoietic cells. J Tiss Cult Meth 13: 55–62, 1991

    Google Scholar 

  21. Emerman JT, Eaves CJ: Lack of effect of hematopoietic growth factors on human breast epithelial cell growth in serum-free primary culture. Bone Marrow Transplant 13: 285–291, 1994

    PubMed  Google Scholar 

  22. Pinkus GS, O'Connor EM, Etheridge CL, Corson JM: Optimal immunoreactivity of keratin proteins in formalin-fixed, paraffin-embedded tissue requires preliminary trypsinization. J Histochem Cytochem 33: 465–473, 1985

    PubMed  Google Scholar 

  23. Latza U, Niedobitek G, Schwarting G, Nekarda H, Stein H: Ber-EP4: new monoclonal antibody which distinguishes epithelia from mesothelia. J Clin Pathol 43: 213–219, 1990

    PubMed  Google Scholar 

  24. Momburg F, Modenhauer G, Hammerling GJ, Moller P: Immunohistochemical study of the expression of a MT 34,000 human epithelium-specific surface glycoprotein in normal and malignant tissues. Cancer Res 47: 2883–2891, 1987

    PubMed  Google Scholar 

  25. Asch BB, Asch HL: Structural components as markers of differentiation and neoplastic progression in mammary epithelial cells. In: Cellular and Molecular Biology of Mammary Cancer. Plenum Press, New York, 1987, pp 29–45

    Google Scholar 

  26. Johnston CS, Shpall EJ, Williams S, Hami L, Jones RB, Bast RC, Ceriani RL, Franklin W: Detection of minimal residual breast cancer in bone marrow. In: Advances in Bone Marrow Purging and Processing. Wiley-Liss, 1992, pp 637–642

  27. Sauvageau C, Lansdorp PM, Eaves CJ, Hogge DE, Dragowska WH, Reid DS, Largman C, Lawrence HJ, Humphries RK: Differential expression of homeobox genes in functionally distinct CD34+ subpopulations of human bone marrow cells. Proc Natl Acad Sci USA 91: 12223–12227, 1994

    PubMed  Google Scholar 

  28. Gabelman BM, Emerman JT: Effects of estrogen, epidermal growth factor, and transforming growth factor-α on the growth of human breast epithelial cells in primary culture. Exp Cell Res 201: 113–118, 1992

    PubMed  Google Scholar 

  29. Sutherland HJ, Eaves CJ, Lansdorp PM, Thacker JD, Hogge DE: Differential regulation of primitive human hematopoietic cells in long-term cultures maintained on genetically engineered murine stromal cells. Blood 78: 666–672, 1991

    PubMed  Google Scholar 

  30. Smith HS, Zoli W, Volpi A, Hiller A, Lippman M, Swain S, Mayall B, Dollbaum C, Hackett AJ, Amadori D: Preliminary correlations of clinical outcome within vitro chemosensitivity of second passage human breast cancer cells. Cancer Res 50: 2943–2948, 1990

    PubMed  Google Scholar 

  31. Zoli W, Volpi A, Bonaguri C, Riccobon A, Savini S, Brizio R, Saragoni A, Medri L, Marra GA, Amadori D: An efficient method of culturing human breast carcinoma to evaluate antiblastic drug activityin vitro: Experience in 136 primary cancers and on 116 recurrences. Breast Cancer Res Treat 17: 231–238, 1990

    Google Scholar 

  32. Bergstraesser LM, Weitzman SA: Culture of normal and malignant primary human mammary epithelial cells in a physiological manner simulatesin vivo growth patterns and allows discrimination of cell type. Cancer Res 53: 2644–2654, 1993

    PubMed  Google Scholar 

  33. Petersen OW, Ronnov-Jessen L, Howlett AR, Bissell MJ: Interaction with basement membrane serves to rapidly distinguish growth and differentiation pattern of normal and malignant human breast epithelial cells. Proc Natl Acad Sci USA 89: 9064–9068, 1992

    PubMed  Google Scholar 

  34. Steel GG: Growth kinetics of tumors: Cell proliferation kinetics in relation to the growth and treatment of cancer. In: Anonymous Oxford Clarendon Press, 1977

  35. Joshi SS, Novak DJ, Messbarger L, Maitreyan V, Weisenburger DD, Sharp JG: Levels of detection of tumor cells in human bone marrow with or without prior culture. Bone Marrow Transplant 6: 179–183, 1990

    PubMed  Google Scholar 

  36. Band V, Sager R: Distinctive traits of normal and tumor-derived human mammary epithelial cells expressed in a medium that supports long-term growth of both cell types. Proc Natl Acad Sci USA 86: 1249–1253, 1989

    PubMed  Google Scholar 

  37. Taylor-Papadimitriou J, Stampfer M, Bartek J, Lewis A, Boshell M, Lane EB, Leigh IM: Keratin expression in human mammary epithelial cells cultured from normal and malignant tissue: Relation toin vivo phenotypes and influence of medium. J Cell Sci 94: 403–413, 1989

    PubMed  Google Scholar 

  38. Hall L, Henney A, Ralphs DNL, Herries DG, Craig RK: Changes in gene expression in established human mammary tumor cell lines when compared with normal breast and breast tumor tissue. Cancer Res 46: 5786–5794, 1986

    PubMed  Google Scholar 

  39. Sharp JG, Armitage J, Crouse D, Joshi S, Kessinger A, Mann S, Vaughan W, Weisenburger D: Recent progress in the detection of metastatic tumor in bone marrow by culture techniques. In: Dicke KA, Spitzer G, Jagannath S, Evinger-Hodges MJ (eds) Autologous Bone Marrow Transplantation. Proceedings of the Fourth International Symposium, The University of Texas M.D. Anderson Cancer Center, Houston, Texas, 1989, pp 421–425

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Anatomy, University of British Columbia, 2177 Wesbrook Mall, V6T 1Z3, Vancouver, B.C., Canada

    Joanne T. Emerman, John Stingl & Andrea Petersen

  2. University of Colorado Hospital, BMT Program, Health Sciences Center, 4200 East 9th Avenue, Box B190, 80262, Denver, Colorado, USA

    Elizabeth J. Shpall

  3. Department of Medical Genetics, University of British Columbia, Canada

    Connie J. Eaves

  4. Terry Fox Laboratory, British Columbia Cancer Agency, Vancouver, B.C., Canada

    Connie J. Eaves

Authors
  1. Joanne T. Emerman
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. John Stingl
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Andrea Petersen
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Elizabeth J. Shpall
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Connie J. Eaves
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Emerman, J.T., Stingl, J., Petersen, A. et al. Selective growth of freshly isolated human breast epithelial cells cultured at low concentrations in the presence or absence of bone marrow cells. Breast Cancer Res Tr 41, 147–159 (1996). https://doi.org/10.1007/BF01807160

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF01807160

Key words

Search

Navigation