MCF-12F细胞
数量： 大量
器官来源： **
细胞形态： 上皮样
生长状态： 贴壁生长
ATCC Number： CRL-10783™
相关**： 正常
年限： 60 years
运输方式： 冻存运输
是否是肿瘤细胞： 0
物种来源： 人
Designations: MCF-12F
Depositors: Michigan Cancer Foundation
MCF-12F细胞Biosafety Level: 1
Shipped: frozen
Medium & Serum: See Propagation
Growth Properties: adherent
Organism: Homo sapiens
Morphology: epithelial
Source: Organ: mammary gland; breast
Disease: normal
Cellular Products: epithelial mucin; sialomucin; milk fat globule membrane antigen
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Tumorigenic: No
Antigen Expression: Blood Type B, RH +
DNA Profile (STR): Amelogenin: X
CSF1PO: 10,11
D13S317: 9,11
D16S539: 9,12
D5S818: 11,13
MCF-12F细胞D7S820: 8,11
THO1: 7
TPOX: 8
vWA: 18
Cytogenetic Analysis: aneuploid
Isoenzymes: AK-1, 1
ES-D, 1
G6PD, B
GLO-I, 1-2
PGM1, 1
PGM3, 1
Age: 60 years
Gender: female
Ethnicity: Caucasian
Comments: The MCF-12F cell line is a non-tumorigenic epithelial cell line established from tissue taken ateduction 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).
MCF-12F细胞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.
Propagation: ATCC complete growth medium: A 1:1 mixture of Dulbecco's modified Eagle's medium and Ham's F12 medium with 0.04 mM Ca++, 20 ng/ml epidermal growth factor, 100 ng/ml cholera toxin, 0.01 mg/ml insulin and 500 ng/ml hydrocortisone, 95%; Chelex-treated horse serum, 5%
Temperature: 37.0°C
Subculturing: Protocol:
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 xg 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
Preservation: Freeze medium: MCF-12F细胞Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Doubling Time: 20 hrs
References: 22025: Pauley RJ, et al. Immortal human mammary epithelial cell sublines. US Patent 5,206,165 dated Apr 27 1993
22864: Paine TM, et al. Characterization of epithelial phenotypes in mortal and immortal human breast cells. Int. J. Cancer 50: 463-473, 1992. PubMed: 1370949
32244: 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
