• 名称: MCF-7(人乳腺癌细胞)
  • 货号: CBP60380
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CBP60380

 

 

I. General information

Synonyms:

MCF 7; MCF.7; MCF7; Michigan Cancer Foundation-7; ssMCF-7; ssMCF7; MCF7/WT; IBMF-7; MCF7-CTRL

Background:

Established from the pleural effusion from a 69 year female caucasian suffering from a breast adenocarcinoma. Cells exhibit some features of differentiated mammary epithelium including oestradiol synthesis and formation of domes. Cells may carry B or C type retrovirus and are considered to represent a category 2 pathogen (P2 containment). Cells express both the wild type and variant oestrogen receptors as well as progesterone receptor. The cells should be handled under laboratory containment level 2.

Species:

Homo sapiens, human

Tissue:

Mammary gland, breast; derived from metastatic site: pleural effusion

Disease:

Breast adenocarcinoma

Gender:

Female, 69 years

Morphology:

Epithelial-like cells growing as monolayers

Growth Mode:

Adherent

Doubling Time:

ca. 50 hours (range 30-72 hours)

DNA Profile:

Amelogenin: X
CSF1PO: 10
D13S317: 11
D16S539: 11,12
D5S818: 11,12/12,12
D7S820: 8,9
THO1: 6
TPOX: 9,12

vWA: 14,15
Cobioer’s Cell Line Authentication Service

Culture Medium:

MEM + 10% FBS+ 1% Non Essential Amino Acids (NEAA) + 1mM Sodium Pyruvate (NaP)+ 10 µg/ml human insulininsulin(optional)
We strongly suggest to purchase the complete medium from Cobioer.

Cryopreservation medium:

90% FBS+10%DMSO

Photo:

 

Antigen Expression:

Blood Type O; Rh+

Receptor Expression:

Estrogen receptor, expressed

Genes Expressed:

Insulin-like growth factor binding proteins (IGFBP) BP-2; BP-4; BP-5

Immunology:

cytokeratin +, cytokeratin-7 -, cytokeratin-8 +, cytokeratin-17 -, cytokeratin-18 +, cytokeratin-19 +, desmin -, endothel -, EpCAM +, GFAP -, neurofilament -, vimentin -

Molec. Genetics:

Expression of fusion gene BCAS4-BCAS3 confirmed by RT-PCR

Viruses:

ELISA: reverse transcriptase negative; PCR: EBV -, HBV -, HCV -, HHV-8 -, HIV -, HTLV-I/II -, MLV -, SMRV -

Comments:

The MCF7 line retains several characteristics of differentiated mammary epithelium including ability to process estradiol via cytoplasmic estrogen receptors and the capability of forming domes. The cells express the WNT7B oncogene.
Growth of MCF7 cells is inhibited by tumor necrosis factor alpha (TNF alpha).

Secretion of IGFBP's can be modulated by treatment with anti-estrogens.

 

II. Handling Procedure for Flask Cultures

The flask was seeded with cells grown and completely filled with medium at Cobioer to prevent loss of cells during shipping.
1.Upon receipt visually examine the culture for macroscopic evidence of any microbial contamination.
Using an inverted microscope (preferably equipped with phasecontrast optics), carefully check for any evidence of microbial contamination. Also check to determine if the majority of cells are still attached to the bottom of the flask? during shipping the cultures are sometimes handled roughly and many of the cells often detach and become suspended in the culture medium (but are still viable).
2.If the cells are still attached, aseptically remove all but 5 to 10 mL of the shipping medium. The shipping medium can be saved for reuse. Incubate the cells at 37°C in a 5% CO2 in air atmosphere until they are ready to be subcultured.
3.If the cells are not attached, aseptically remove the entire contents of the flask and centrifuge at 1000rpm for 5 to 10 minutes. Remove shipping medium and save. Resuspend the pelleted cells in 10 mL of this medium and add to 25 cm2 flask. Incubate at 37°C in a 5% CO2 in air atmosphere until cells are ready to be subcultured.

 

III. Subculturing Procedure

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) Trypsin0.53 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 (depend on the batch of trypsin).
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.Add appropriate aliquots of the cell suspension to new culture vessels.
6.Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:6 is recommended
Medium Renewal: 2 to 3 times per week

 

IV. Cryopreservation Procedure

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) Trypsin0.53mM 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 within 1 to 10 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 a centrifuge tube and spin at approximately 1000rpmfor 5 to10 minutes.
6.Discard supernatant and resuspend cells in cryopreservation medium.
7.Transfer the cells into Cryogenic Vials, 1ml/vial.
8.Frozen the cells in cryogenic container (Nalgene #5100-0001).

 

V. Database

Mutation:

Mutation Gene

Zygosity

Gene Sequence

Protein Sequence

CDKN2A

Homozygous

c.1_471del471

p.0?

PIK3CA

Heterozygous

c.1633G>A

p.E545K

ERBB4

Unknown

c.3725A>G

p.Y1242C

For more information, Please visit
http://www.cobioer.com/sjk/&pmcId=77.html

Expression:

GSG1L, TRPM7, USP8
For more information, Please visit
http://www.cobioer.com/sjk/&pmcId=77.html

Comments:

For more information, please contact Cobioer(4008-750-250).

 

VI. References

1.PubMed=23933261; DOI=10.1016/j.celrep.2013.07.018; Moghaddas Gholami A., Hahne H., Wu Z., Auer F.J., Meng C., Wilhelm M., Kuster B.; "Global proteome analysis of the NCI-60 cell line panel."; Cell Rep. 4:609-620(2013).

2.PubMed=25321415; DOI=10.1210/me.2014-1229; Li Y., Arao Y., Hall J.M., Burkett S., Liu L., Gerrish K., Cavailles V., Korach K.S.; "Research resource: STR DNA profile and gene expression comparisons of human BG-1 cells and a BG-1/MCF-7 clonal variant."; Mol. Endocrinol. 28:2072-2081(2014).

3.PubMed=25485619; DOI=10.1038/nbt.3080; Klijn C., Durinck S., Stawiski E.W., Haverty P.M., Jiang Z., Liu H., Degenhardt J., Mayba O., Gnad F., Liu J., Pau G., Reeder J., Cao Y., Mukhyala K., Selvaraj S.K., Yu M., Zynda G.J., Brauer M.J., Wu T.D., Gentleman R.C., Manning G., Yauch R.L., Bourgon R., Stokoe D., Modrusan Z., Neve R.M., de Sauvage F.J., Settleman J., Seshagiri S., Zhang Z.; "A comprehensive transcriptional portrait of human cancer cell lines."; Nat. Biotechnol. 33:306-312(2015).

4.PubMed=26055192; DOI=10.1021/acs.jproteome.5b00375; Cifani P., Kirik U., Waldemarson S., James P.; "Molecular portrait of breast-cancer-derived cell lines reveals poor similarity with tumors."; J. Proteome Res. 14:2819-2827(2015).
5.PubMed=26330541; DOI=10.1074/mcp.M115.050484; Wu X., Zahari M.S., Renuse S., Nirujogi R.S., Kim M.-S., Manda S.S., Stearns V., Gabrielson E., Sukumar S., Pandey A.; "Phosphoproteomic analysis identifies focal adhesion kinase 2 (FAK2) as a potential therapeutic target for tamoxifen resistance in breast cancer."; Mol. Cell. Proteomics 14:2887-2900(2015).
6.PubMed=27362937; DOI=10.1371/journal.pone.0157290; Carrier M., Joint M., Lutzing R., Page A., Rochette-Egly C.; "Phosphoproteome and transcriptome of RA-responsive and RA-resistant breast cancer cell lines."; PLoS ONE 11:E0157290-E0157290(2016).
7.PubMed=27397505; DOI=10.1016/j.cell.2016.06.017; Iorio F., Knijnenburg T.A., Vis D.J., Bignell G.R., Menden M.P., Schubert M., Aben N., Goncalves E., Barthorpe S., Lightfoot H., Cokelaer T., Greninger P., van Dyk E., Chang H., de Silva H., Heyn H., Deng X., Egan R.K., Liu Q., Mironenko T., Mitropoulos X., Richardson L., Wang J., Zhang T., Moran S., Sayols S., Soleimani M., Tamborero D., Lopez-Bigas N., Ross-Macdonald P., Esteller M., Gray N.S., Haber D.A., Stratton M.R., Benes C.H., Wessels L.F.A., Saez-Rodriguez J., McDermott U., Garnett M.J.; "A landscape of pharmacogenomic interactions in cancer."; Cell 166:740-754(2016).
8.PubMed=27456714; DOI=10.1038/srep28994; Kleensang A., Vantangoli M.M., Odwin-DaCosta S., Andersen M.E., Boekelheide K., Bouhifd M., Fornace A.J. Jr., Li H.-H., Livi C.B., Madnick S., Maertens A., Rosenberg M., Yager J.D., Zhaog L., Hartung T.; "Genetic variability in a frozen batch of MCF-7 cells invisible in routine authentication affecting cell function."; Sci. Rep. 6:28994-28994(2016).

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