| 產(chǎn)品名稱 |
MCF-12F |
| 商品貨號 |
B165112 |
| Organism |
Homo sapiens, human |
| Tissue |
mammary gland; breast |
| Cell Type |
Epithelial |
| Product Format |
frozen |
| Morphology |
epithelial |
| Culture Properties |
adherent |
| Biosafety Level |
1
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. |
| Disease |
normal |
| Age |
60 years |
| Gender |
female |
| Ethnicity |
Caucasian |
| Storage Conditions |
liquid nitrogen vapor phase |
| Disclosure |
This material is cited in a US or other Patent and may not be used to
infringe the claims. Depending on the wishes of the Depositor, ATCC may be
required to inform the Patent Depositor of the party to which the material was
furnished. This material may not have been produced or characterized by ATCC. |
| Karyotype |
aneuploid |
| Derivation |
The MCF-12F cell line is a non-tumorigenic epithelial cell line established from tissue taken at reduction mammoplasty from a nulliparous patient with fibrocystic breast disease that contained focal areas of intraductal hyperplasia.
The line was produced by long term culture in serum free medium with low Ca++ concentration.
MCF-12F was derived from floating cells in the population.
Cells derived from an adherent population are available (see MCF-12A, ATCC CRL-10782). |
| Antigen Expression |
Blood Type B, RH + |
| Genes Expressed |
epithelial mucin; sialomucin; milk fat globule membrane antigen |
| Cellular Products |
epithelial mucin; sialomucin; milk fat globule membrane antigen |
| Tumorigenic |
No |
| Effects |
No, in immunosuppressed mice Yes, in semisolid medium |
| Comments |
The cells are positive for epithelial cytokeratins 8, 14 and 18, and negative for cytokeratin 19.
They exhibit typical luminal epithelial morphology, three dimensional growth in collagen, and form domes in confluent cultures. |
| Complete Growth Medium |
Base medium: Combine 14.8g/L Dulbecco's modified Eagle's medium and Ham's F12 base (Sigma, D-9785), 1.2g NaHCO3 (Sigma, S-5761), 0.365g L-glutamine (Sigma, G-3126), 0.059g L-leucine (Sigma, L-8912), 0.0912g L-lysine (Sigma, L-8662), 0.017g L-methionine (Sigma, M-5308), 0.0612g MgCl2.6H2O (Sigma, M-1028), 0.0488g MgSO4.7H2O (Sigma, M-3409), 0.006g CaCl2.2H2O (Sigma, C-8106), and 0.0086g Phenol Red (Sigma, P-3532). Fill to 1L with Ultrapure Cell Grade water (ATCC? 30-2205). Stir to dissolve. Adjust pH to 7.1 – 7.3. Filter-sterilize using a 0.22 μm filter. Complete growth medium: Combine base medium with 20 ng/mL epidermal growth factor (Sigma, E-9644), 100 ng/mL cholera toxin (Sigma, C-8052), 0.01 mg/mL human insulin (Sigma, I-2643), 500 ng/mL hydrocortisone (Sigma, H-0888), and 5% Chelex-treated horse serum.
|
| Subculturing |
Volumes used in this protocol are for 75 cm2 flask; 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 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that 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.
- To remove trypsin-EDTA solution, transfer cell suspension to centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes. Discard supernatant and resuspend cells in fresh growth medium. Add appropriate aliquots of cell suspension to new culture vessels.
- Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:2 is recommended
Medium Renewal: Every 2 to 3 days
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994. |
| Cryopreservation |
Complete growth medium supplemented with 5% (v/v) DMSO. Cell culture tested DMSO is available as ATCC Catalog No. 4-X. |
| Culture Conditions |
Temperature: 37°C |
| STR Profile |
Amelogenin: X CSF1PO: 10,11 D13S317: 9,11 D16S539: 9,12 D5S818: 11,13 D7S820: 8,11 THO1: 7 TPOX: 8 vWA: 18 |
| Isoenzymes |
AK-1, 1 ES-D, 1 G6PD, B GLO-I, 1-2 PGM1, 1 PGM3, 1 |
| Population Doubling Time |
20 hrs |
| Name of Depositor |
Michigan Cancer Foundation |
| Deposited As |
Homo sapiens |
| U.S. Patent Number |
|
| References |
Pauley RJ, et al. Immortal human mammary epithelial cell sublines. US Patent 5,206,165 dated Apr 27 1993
Paine TM, et al. Characterization of epithelial phenotypes in mortal and immortal human breast cells. Int. J. Cancer 50: 463-473, 1992. PubMed: 1370949
Hoppe HC, et al. Identification of phosphatidylinositol mannoside as a mycobacterial adhesin mediating both direct and opsonic binding to nonphagocytic mammalian cells. Infect. Immun. 65: 3896-3905, 1997. PubMed: 9284169
Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.
Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.
Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.
Biosafety in Microbiological and Biomedical Laboratories, 5th ed. HHS. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 2007. The entire text is available online.
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