BCRJ Code | 0162 |
Cell Line | MCF7 |
Species | Homo sapiens |
Vulgar Name | Human |
Tissue | Mammary Gland, Breast; Derived From Metastatic Site: Pleural Effusion |
Cell Type | Epithelial |
Morphology | Epithelial |
Disease | Adenocarcinoma |
Growth Properties | Adherent |
Sex | Female |
Age/Ethinicity | 69 Year / |
Applications | These cells are a suitable transfection host |
DNA Profile | Amelogenin: X CSF1PO: 10 D13S317: 11 D16S539: 11,12 D5S818: 11,12 D7S820: 8,9 THO1: 6 TPOX: 9,12 vWA: 14,15 |
Products | insulin-like growth factor binding proteins (IGFBP) BP-2; BP-4; BP-5 |
Biosafety | 1 |
Addtional Info | The MCF7 line retains several characteristics of differentiated mammary epithelium including ability to process estradiol via cytoplasmic estrogen receptors and the capability of forming domes. The cells express the WNT7B oncogene. [PubMed: 8168088] Growth of MCF7 cells is inhibited by tumor necrosis factor alpha (TNF alpha). Secretion of IGFBP's can be modulated by treatment with anti-estrogens. |
Culture Medium | RPMI-1640 medium modified to contain 2 mM L-glutamine, 1 mM sodium pyruvate, 4500 mg/L glucose and 20% of fetal bovine serum. |
Subculturing | Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. Remove and discard culture medium. Briefly rinse the cell layer with PBS without calcium and magnesium to remove all traces of serum which contains trypsin inhibitor. Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes). Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting. Add appropriate aliquots of the cell suspension to new culture vessels. Incubate cultures at 37°C. Population Doubling Time approximately 29 hours NOTE: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 12 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 6th edition, published by Alan R. Liss, N.Y., 2010. |
Subculturing Medium Renewal | 2 to 3 times per week |
Subculturing Subcultivation Ratio | 1:3 to 1:6 |
Culture Conditions | Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37°C |
Cryopreservation | 95% FBS + 5% DMSO (Dimethyl sulfoxide) |
Thawing Frozen Cells | SAFETY PRECAUTION:
It is strongly recommended to always wear protective gloves, clothing, and a full-face mask when handling frozen vials. Some vials may leak when submerged in liquid nitrogen, allowing nitrogen to slowly enter the vial. Upon thawing, the conversion of liquid nitrogen back to its gas phase may cause the vial to explode or eject its cap with significant force, creating flying debris.
NOTE: It is important to avoid excessive alkalinity of the medium during cell recovery. To minimize this risk, it is recommended to place the culture vessel containing the growth medium in the incubator for at least 15 minutes before adding the vial contents. This allows the medium to stabilize at its normal pH (7.0 to 7.6). |
References | Sugarman BJ, et al. Recombinant human tumor necrosis factor-alpha: effects on proliferation of normal and transformed cells in vitro. Science 230: 943-945, 1985. PubMed: 3933111 Takahashi K, Suzuki K. Association of insulin-like growth-factor-I-induced DNA synthesis with phosphorylation and nuclear exclusion of p53 in human breast cancer MCF-7 cells. Int. J. Cancer 55: 453-458, 1993. PubMed: 8375929 Brandes LJ, Hermonat MW. Receptor status and subsequent sensitivity of subclones of MCF-7 human breast cancer cells surviving exposure to diethylstilbestrol. Cancer Res. 43: 2831-2835, 1983. PubMed: 6850594 Lan MS, et al. Polypeptide core of a human pancreatic tumor mucin antigen. Cancer Res. 50: 2997-3001, 1990. PubMed: 2334903 Pratt SE, Pollak MN. Estrogen and antiestrogen modulation of MCF7 human breast cancer cell proliferation is associated with specific alterations in accumulation of insulin-like growth factor-binding proteins in conditioned media. Cancer Res. 53: 5193-5198, 1993. PubMed: 7693333 Huguet EL, et al. Differential expression of human Wnt genes 2, 3, 4, and 7B in human breast cell lines and normal and disease states of human breast tissue. Cancer Res. 54: 2615-2621, 1994. PubMed: 8168088 Soule HD, et al. A human cell line from a pleural effusion derived from a breast carcinoma. J. Natl. Cancer Inst. 51: 1409-1416, 1973. PubMed: 4357757 Bellet D, et al. Malignant transformation of nontrophoblastic cells is associated with the expression of chorionic gonadotropin beta genes normally transcribed in trophoblastic cells. Cancer Res. 57: 516-523, 1997. PubMed: 9012484 Littlewood-Evans AJ, et al. The osteoclast-associated protease cathepsin K is expressed in human breast carcinoma. Cancer Res. 57: 5386-5390, 1997. PubMed: 9393764 Komarova EA, et al. Intracellular localization of p53 tumor suppressor protein in gamma-irradiated cells is cell cycle regulated and determined by the nucleus. Cancer Res. 57: 5217-5220, 1997. PubMed: 9393737 van Dijk MA, et al. A functional assay in yeas for the human estrogen receptor displays wild-type and variant estrogen receptor messenger RNAs present in breast carcinoma. Cancer Res. 57: 3478-3485, 1997. PubMed: 9270016 Landers JE, et al. Translational enhancement of mdm2 oncogene expression in human tumor cells containing a stabilized wild-type p53 protein. Cancer Res. 57: 3562-3568, 1997. PubMed: 9270029 Umekita Y, et al. Human prostate tumor growth in athymic mice: inhibition by androgens and stimulation by finasteride. Proc. Natl. Acad. Sci. USA 93: 11802-11807, 1996. PubMed: 8876218 Zamora-Leon SP, et al. Expression of the fructose transporter GLUT5 in human breast cancer. Proc. Natl. Acad. Sci. USA 93: 1847-1852, 1996. PubMed: 8700847 Geiger T, et al. Antitumor activity of a PKC-alpha antisense oligonucleotide in combination with standard chemotherapeutic agents against various human tumors transplanted into nude mice. Anticancer Drug Des. 13: 35-45, 1998. PubMed: 9474241 Jang SI, et al. Activator protein 1 activity is involved in the regulation of the cell type-specific expression from the proximal promoter of the human profilaggrin gene. J. Biol. Chem. 271: 24105-24114, 1996. PubMed: 8798649 Lee JH, et al. The proximal promoter of the human transglutaminase 3 gene. J. Biol. Chem. 271: 4561-4568, 1996. PubMed: 8626812 Chang K, Pastan I. Molecular cloning of mesothelin, a differentiation antigen present on mesothelium, mesotheliomas, and ovarian cancers. Proc. Natl. Acad. Sci. USA 93: 136-140, 1996. PubMed: 8552591 Zhu X, et al. Cell cycle-dependent modulation of telomerase activity in tumor cells. Proc. Natl. Acad. Sci. USA 93: 6091-6095, 1996. PubMed: 8650224 Bacus SS, et al. Differentiation of cultured human breast cancer cells (AU-565 and MCF-7) associated with loss of cell surface HER-2/neu antigen. Mol. Carcinog. 3: 350-362, 1990. PubMed: 1980588 Huguet EL, et al. Differential expression of human Wnt genes 2, 3, 4, and 7B in human breast cell lines and normal and disease states of human breast tissue. Cancer Res. 54: 2615-2621, 1994. PubMed: 8168088 |
Depositors | Ricardo Bretani; Ludwick Institute For Cancer Research São Paulo - Sp |
ATCC | HTB-22 |
Cellosaurus | CVCL_0031 |
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