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Isolation and Characterization of a Spontaneously Immortalized Human Breast Epithelial Cell Line, MCF-10¹

Michigan Cancer Foundation [H. D. S., T. M. M., S. R. W., R. B., J. R., R. J. P., R. F. J., S. C. B.] and Departments of Pediatrics [W. D. P.] and Biochemistry [S. C. B.], Wayne State University School of Medicine, Children's Hospital of Michigan, Detroit, Michigan 48201

Two sublines of a breast epithelial cell culture, MCF-10, derived from human fibrocystic mammary tissue exhibit immortality after extended cultivation in low calcium concentrations (0.03–0.06 mM) and floating transfers in low calcium (MCF-10F), or by trypsin-Versene passages in the customary (normal) calcium levels, 1.05 mM (MCF-10A). Both sublines have been maintained as separate entities after 2.3 years (849 days) in vitro and at present have been in culture for longer than 4 years. MCF-10 has the characteristics of normal breast epithelium by the following criteria: (a) lack of tumorigenicity in nude mice; (b) three-dimensional growth in collagen; (c) growth in culture that is controlled by hormones and growth factors; (d) lack of anchorage-independent growth; and (e) dome formation in confluent cultures. Cytogenetic analysis prior to immortalization showed normal diploid cells; although later passages showed minimal rearrangement and near-diploidy, the immortal cells were not karyotypically normal. The emergence of an immortal culture in normal calcium media was not an inherent characteristic of the original tissue from which MCF-10 was derived since reactivated cryopreserved cells from cultures grown for 0.3 and 1.2 years in low calcium were incapable of sustained growth in normal calcium.

¹ These investigations were supported in part by NIH Grants CA28999, CA36399, and CA38921 and institutional grants to the Michigan Cancer Foundation from the United Foundation of Greater Detroit and from Abbott Laboratories. Cell characterization studies were carried out under National Cancer Institute Contract N01-CP-21017 to Children's Hospital of Michigan.

Received 11/27/89. Accepted 6/12/90.

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				B. R. Baldwin, N. A. Timchenko, and C. A. Zahnow Epidermal Growth Factor Receptor Stimulation Activates the RNA Binding Protein CUG-BP1 and Increases Expression of C/EBP{beta}-LIP in Mammary Epithelial Cells Mol. Cell. Biol., May 1, 2004; 24(9): 3682 - 3691. [Abstract] [Full Text] [PDF]

				S. E. Seton-Rogers, Y. Lu, L. M. Hines, M. Koundinya, J. LaBaer, S. K. Muthuswamy, and J. S. Brugge Cooperation of the ErbB2 receptor and transforming growth factor {beta} in induction of migration and invasion in mammary epithelial cells PNAS, February 3, 2004; 101(5): 1257 - 1262. [Abstract] [Full Text] [PDF]

				Y. Hoshiya, V. Gupta, H. Kawakubo, E. Brachtel, J. L. Carey, L. Sasur, A. Scott, P. K. Donahoe, and S. Maheswaran Mullerian Inhibiting Substance Promotes Interferon {gamma}-induced Gene Expression and Apoptosis in Breast Cancer Cells J. Biol. Chem., December 19, 2003; 278(51): 51703 - 51712. [Abstract] [Full Text] [PDF]

				F. Tian, S. DaCosta Byfield, W. T. Parks, S. Yoo, A. Felici, B. Tang, E. Piek, L. M. Wakefield, and A. B. Roberts Reduction in Smad2/3 Signaling Enhances Tumorigenesis but Suppresses Metastasis of Breast Cancer Cell Lines Cancer Res., December 1, 2003; 63(23): 8284 - 8292. [Abstract] [Full Text] [PDF]

				J. Debnath, S. J. Walker, and J. S. Brugge Akt activation disrupts mammary acinar architecture and enhances proliferation in an mTOR-dependent manner J. Cell Biol., October 27, 2003; 163(2): 315 - 326. [Abstract] [Full Text] [PDF]

				S. L. Starcevic, N. M. Diotte, K. L. Zukowski, M. J. Cameron, and R. F. Novak Oxidative DNA Damage and Repair in a Cell Lineage Model of Human Proliferative Breast Disease (PBD) Toxicol. Sci., September 1, 2003; 75(1): 74 - 81. [Abstract] [Full Text] [PDF]

				J. Lai, J. Chien, J. Staub, R. Avula, E. L. Greene, T. A. Matthews, D. I. Smith, S. H. Kaufmann, L. R. Roberts**, and V. Shridhar** Loss of HSulf-1 Up-regulates Heparin-binding Growth Factor Signaling in Cancer J. Biol. Chem., June 13, 2003; 278(25): 23107 - 23117. [Abstract] [Full Text] [PDF]

				W. Yu, B. G. Sanders, and K. Kline RRR-{alpha}-Tocopheryl Succinate-induced Apoptosis of Human Breast Cancer Cells Involves Bax Translocation to Mitochondria Cancer Res., May 15, 2003; 63(10): 2483 - 2491. [Abstract] [Full Text] [PDF]

				P. K. Sengupta, E. M. Smith, K. Kim, M. J. Murnane, and B. D. Smith DNA Hypermethylation Near the Transcription Start Site of Collagen {alpha}2(I) Gene Occurs in Both Cancer Cell Lines and Primary Colorectal Cancers Cancer Res., April 15, 2003; 63(8): 1789 - 1797. [Abstract] [Full Text] [PDF]

				A. Lyakhovich and M. P. V. Shekhar Supramolecular Complex Formation between Rad6 and Proteins of the p53 Pathway during DNA Damage-Induced Response Mol. Cell. Biol., April 1, 2003; 23(7): 2463 - 2475. [Abstract] [Full Text] [PDF]

				S. O. Mueller, J. M. Hall, D. L. Swope, L. C. Pedersen, and K. S. Korach Molecular Determinants of the Stereoselectivity of Agonist Activity of Estrogen Receptors (ER) alpha and beta J. Biol. Chem., March 28, 2003; 278(14): 12255 - 12262. [Abstract] [Full Text] [PDF]

				X. Zhang, T. Zhu, Y. Chen, H. C. Mertani, K.-O. Lee, and P. E. Lobie Human Growth Hormone-regulated HOXA1 Is a Human Mammary Epithelial Oncogene J. Biol. Chem., February 21, 2003; 278(9): 7580 - 7590. [Abstract] [Full Text] [PDF]

				J. L. Martin, S. M. Weenink, and R. C. Baxter Insulin-like Growth Factor-binding Protein-3 Potentiates Epidermal Growth Factor Action in MCF-10A Mammary Epithelial Cells. INVOLVEMENT OF p44/42 AND p38 MITOGEN-ACTIVATED PROTEIN KINASES J. Biol. Chem., January 24, 2003; 278(5): 2969 - 2976. [Abstract] [Full Text] [PDF]

				R. S. Frey and K. W. Singletary Genistein Activates p38 Mitogen-Activated Protein Kinase, Inactivates ERK1/ERK2 and Decreases Cdc25C Expression in Immortalized Human Mammary Epithelial Cells J. Nutr., January 1, 2003; 133(1): 226 - 231. [Abstract] [Full Text] [PDF]

				Z. Li, Z. H. Meng, A. Sayeed, R. Shalaby, B.-M. Ljung, and S. H. Dairkee Genome-wide Allelotyping of a New in Vitro Model System Reveals Early Events in Breast Cancer Progression Cancer Res., October 15, 2002; 62(20): 5980 - 5987. [Abstract] [Full Text] [PDF]

				Y. Kodama, R. C. Baxter, and J. L. Martin Insulin-Like Growth Factor-I Inhibits Cell Growth in the A549 Non-Small Lung Cancer Cell Line Am. J. Respir. Cell Mol. Biol., September 1, 2002; 27(3): 336 - 344. [Abstract] [Full Text] [PDF]

				A. Ryo, Y.-C. Liou, G. Wulf, M. Nakamura, S. W. Lee, and K. P. Lu PIN1 Is an E2F Target Gene Essential for Neu/Ras-Induced Transformation of Mammary Epithelial Cells Mol. Cell. Biol., August 1, 2002; 22(15): 5281 - 5295. [Abstract] [Full Text] [PDF]

				J. Botos, R. Smith III, and D. T. Kochevar Retinoblastoma Function Is a Better Indicator of Cellular Phenotype in Cultured Breast Adenocarcinoma Cells than Retinoblastoma Expression Experimental Biology and Medicine, May 1, 2002; 227(5): 354 - 362. [Abstract] [Full Text] [PDF]

				M. Guo, P. A. Mathieu, B. Linebaugh, B. F. Sloane, and J. J. Reiners Jr. Phorbol Ester Activation of a Proteolytic Cascade Capable of Activating Latent Transforming Growth Factor-beta . A PROCESS INITIATED BY THE EXOCYTOSIS OF CATHEPSIN B J. Biol. Chem., April 19, 2002; 277(17): 14829 - 14837. [Abstract] [Full Text] [PDF]

				M. P. V. Shekhar, A. Lyakhovich, D. W. Visscher, H. Heng, and N. Kondrat Rad6 Overexpression Induces Multinucleation, Centrosome Amplification, Abnormal Mitosis, Aneuploidy, and Transformation Cancer Res., April 1, 2002; 62(7): 2115 - 2124. [Abstract] [Full Text] [PDF]

				J.-H. Sheen and R. B. Dickson Overexpression of c-Myc Alters G1/S Arrest following Ionizing Radiation Mol. Cell. Biol., March 15, 2002; 22(6): 1819 - 1833. [Abstract] [Full Text] [PDF]

				J. M. Shields, K. Rogers-Graham, and C. J. Der Loss of Transgelin in Breast and Colon Tumors and in RIE-1 Cells by Ras Deregulation of Gene Expression through Raf-independent Pathways J. Biol. Chem., March 15, 2002; 277(12): 9790 - 9799. [Abstract] [Full Text] [PDF]

				M. Fan, X. Long, J. A. Bailey, C. A. Reed, E. Osborne, E. A. Gize, E. A. Kirk, R. M. Bigsby, and K. P. Nephew The Activating Enzyme of NEDD8 Inhibits Steroid Receptor Function Mol. Endocrinol., February 1, 2002; 16(2): 315 - 330. [Abstract] [Full Text] [PDF]