| 產(chǎn)品名稱 |
MCF 10F |
| 商品貨號(hào) |
B165109 |
| Organism |
Homo sapiens, human |
| Tissue |
mammary gland; breast |
| Cell Type |
Epithelial, Myoepithelial |
| 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 |
fibrocystic disease |
| Age |
36 years |
| Gender |
female |
| 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 human female; 46, XX, 1p+, t(3:9)(p13:p22) |
| Derivation |
Cells derived from an adherent population are available (see MCF 10A, ATCC CRL-10317). MCF 10F was derived from floating cells in the population. The line was produced by long term culture in serum free medium with low Ca++ concentration. |
| Clinical Data |
female
36 years |
| Receptor Expression |
epidermal growth factor (EGF), expressed insulin, expressed glucocorticoid, expressed |
| Tumorigenic |
No |
| Effects |
No, in immunosuppressed mice Yes, in semisolid medium |
| Comments |
The MCF 10F cell line is a non-tumorigenic epithelial cell line.
The cells are positive for epithelial sialomucins.
Cytokeratins and milk fat globule antigen.
They exhibit three dimensional growth in collagen, and form domes in confluent cultures.
Thus far, the cells have shown no signs of terminal differentiation or senescence.
The line is responsive to insulin, glucocorticoids, cholera enterotoxin, and epidermal growth factor (EGF).
By electron microscopy the cells display characteristics of luminal ductal cells but not of myoepithelial cells.
They also express breast specific antigens as detected by positive reaction with MFA-Breast and MC-5 monoclonal antibodies.
The calcium content of the medium exerts a strong effect on the morphology of the cells.
|
| 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 are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally 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 to 1:4 is recommended
Medium Renewal: Every 2 to 3 days |
| Cryopreservation |
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO Storage temperature: liquid nitrogen vapor phase |
| Culture Conditions |
Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37°C |
| STR Profile |
Amelogenin: X CSF1PO: 10,12 D13S317: 8,9 D16S539: 11,12 D5S818: 10,13 D7S820: 10,11 THO1: 8,9.3 TPOX: 9,11 vWA: 15,17 |
| Isoenzymes |
AK-1, 1 ES-D, 1 G6PD, B GLO-I, 1-2 PGM1, 1-2 PGM3, 1 |
| Name of Depositor |
Michigan Cancer Foundation |
| Deposited As |
Homo sapiens |
| U.S. Patent Number |
|
| References |
Soule H, McGrath CM. Immortal human mammary epithelial cell lines. US Patent 5,026,637 dated Jun 25 1991
Pauley RJ, et al. Immortal human mammary epithelial cell sublines. US Patent 5,206,165 dated Apr 27 1993
Soule HD, McGrath CM. A simplified method for passage and long-term growth of human mammary epithelial cells. In Vitro Cell. Dev. Biol. 22: 6-12, 1986. PubMed: 2418007
Soule HD, et al. Isolation and characterization of a spontaneously immortalized human breast epithelial cell line, MCF-10. Cancer Res. 50: 6075-6086, 1990. PubMed: 1975513
Tait L, et al. Ultrastructural and immunocytochemical characterization of an immortalized human breast epithelial cell line, MCF-10. Cancer Res. 50: 6087-6094, 1990. PubMed: 1697506
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