MCF 10F (ATCC® CRL-10318)

Organism: Homo sapiens, human  /  Cell Type: Epithelial, Myoepithelial  /  Tissue: mammary gland; breast  /  Disease: fibrocystic disease

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Organism Homo sapiens, human
Tissue mammary gland; breast
Cell Type Epithelial, Myoepithelial
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 1
Disease fibrocystic disease
Age 36 years
Gender female
Storage Conditions liquid nitrogen vapor phase
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.
  1. Remove and discard culture medium.
  2. 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.
  3. 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.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  5. 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.
  6. 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
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

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
FAQ's
  1. Protocol for making Chelex-treated serum


    Date Updated: 4/4/2014

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