Selective growth of freshly isolated human breast epithelial cells cultured at low concentrations in the presence or absence of bone marrow cells
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.
Key wordshuman breast epithelial cell colonies bone marrow metastatic infiltrates primary culture immunocytochemistry cytokeratins epithelial-specific antigens
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- 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 701–718Google 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, 1993PubMedGoogle 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, 1993PubMedGoogle 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 226–248Google 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, 1993PubMedGoogle 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 56–64Google Scholar
- 20.Eaves CJ, Cashman JD, Eaves AC: Methodology of longterm culture of human hemopoietic cells. J Tiss Cult Meth 13: 55–62, 1991Google 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 29–45Google 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 637–642Google 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, 1990Google 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, 1977Google 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 421–425Google Scholar