| 產品名稱 |
MCF-12A |
| 商品貨號 |
B165111 |
| Organism |
Homo sapiens, human |
| Tissue |
mammary gland; breast |
| Cell Type |
epithelial spontanous immortalization |
| 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. |
| Age |
60 years adult |
| 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 |
Chromosome number range = 65 to 71
(Pauley RJ, et al. Immortal human mammary epithelial cell sublines. US Patent 5,206,165 dated Apr 27 1993) |
| Derivation |
The MCF-12A 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-12A was derived from adherent cells in the population. |
| Clinical Data |
60 years adult Caucasian female |
| Genes Expressed |
epithelial mucin, milk fat globule membrane antigen, sialomucin |
| Cellular Products |
epithelial mucin milk fat globule membrane antigen sialomucin |
| Tumorigenic |
No |
| Effects |
No, in immunosuppressed mice Yes, in semisolid medium. |
| Comments |
MCF-12A cells exhibit typical luminal epithelial morphology, three dimensional growth in collagen, and form domes in confluent cultures.
The cells are positive for epithelial cytokeratins 8, 14 and 18, and negative for cytokeratin 19.
Cells derived from a floating population are available (see MCF-12F, ATCC CRL-10783).
|
| Complete Growth Medium |
A 1:1 mixture of Dulbecco's modified Eagle's medium and Ham's F12 medium, 20 ng/ml Human epidermal growth factor, 100 ng/ml cholera toxin, 0.01 mg/ml bovine insulin and 500 ng/ml hydrocortisone, 95%; horse serum, 5%
|
| 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-0.03% (w/v) EDTA solution 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 with 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 culture medium. Add appropriate aliquots of cell suspension to new culture vessels.
-
Place culture vessels in incubators at 37°C.
Subcultivation inoculum: 1-2 x 104 viable cells/cm2
Medium Renewal: Every 2 to 3 days |
| Cryopreservation |
Freeze Medium: Complete growth medium 95%; DMSO, 5%
Storage Temperature: Liquid nitrogen vapor phase |
| Culture Conditions |
Atmosphere: Air, 95%; carbon dioxide (CO2), 5%
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 |
19 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
Pauley RJ, et al. Immortal human mammary epithelial cell sublines. US Patent 5,206,165 dated Apr 27 1993
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