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MCF-12A (ATCC® CRL-10782™)

Organism: Homo sapiens, human  /  Cell Type: epithelial spontanous immortalization  /  Tissue: mammary gland; breast  / 

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.

  1. Remove and discard culture medium.
  2. 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.
  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 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.
  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.
  6. Discard supernatant and resuspend cells in fresh culture medium.  Add appropriate aliquots of cell suspension to new culture vessels. 
  7. 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
U.S. Patent Number
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.
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

Basic Documentation
Product Sheet
Product Sheet
Certificate of Analysis
Certificate of Analysis
SDS
SDS
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