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MCF-12A

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價格:

編號:B165111

品牌:Mingzhoubio

標準菌株

定量菌液

DNA

RNA

規格:

凍干粉

斜面

甘油

平板

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產品名稱

MCF-12A

商品貨號

B165111

Organism

Homo sapiens, human

Tissue

mammary gland; breast

培養基

Cell Type

epithelial spontanous immortalization

Product Format

frozen

Morphology

epithelial

Culture Properties

adherent

Biosafety Level

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

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 10⁴viable 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

5,206,165

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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