Product sheet info

Product Information Sheet for ATCC® HTB-22™
Cell Line Designation: MCF-7
No DM were detected. Chromosome 20 was nullisomic and X was disomic. ATCC® Catalog No. HTB-22™
Note: Cytogenetic information is based on initial seed stock
at ATCC. Cytogenetic instability has been reported in the Table of Contents:
Purified DNA from this line is available as ATCC® HTB-
Total RNA from this line is available as ATCC® HTB-22R™
• Handling Procedure for Flask Cultures Biosafety Level: 1
Appropriate safety procedures should always be used with this material. Laboratory safety is discussed in the following publication: Biosafety in Microbiological and Biomedical Laboratories, 4th ed. HHS Publication No. (CDC) 93-8395. U.S. Department of Health and Human Services, Centers for Disease Control and Prevention. Washington DC: U.S. Government Printing Office; 1999. The entire text is available Cell Line Description
online at www.cdc.gov/od/ohs/biosfty/bmbl4/bmbl4toc.htm. Organism: Homo sapiens (human)
Tissue: mammary gland; breast adenocarcinoma; derived
Use Restrictions
These cells are distributed for research purposes only.
Age: 69 years
ATCC recommends that individuals contemplating Gender: female
commercial use of any cell line first contact the originating Ethnicity: Caucasian
investigator to negotiate an agreement. Third party Morphology: epithelial
distribution of this cell line is discouraged, since this practice Doubling time: about 29 hours
has resulted in the unintentional spreading of cell lines Growth Properties: adherent
contaminated with inappropriate animal cells or microbes. Oncogene: wnt7h +
Antigens Expressed: Blood Type O; Rh+
Handling Procedure for Frozen Cells
Products: insulin-like growth factor binding proteins
(IGFBP) BP-2; BP-4; BP-5
To insure the highest level of viability, thaw the vial and DNA profile (STR analysis)
initiate the culture as soon as possible upon receipt. If upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at –70°C. Storage at –70°C will result in loss of viability. SAFETY PRECAUTION: ATCC highly recommends that
protective gloves and clothing always be used and a full
face mask always be worn when handling frozen vials. It
is important to note that some vials leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon Depositor: C.M. McGrath
thawing, the conversion of the liquid nitrogen back to its gas Comments: The MCF7 line retains several characteristics of
phase may result in the vessel exploding or blowing off its differentiated mammary epithelium including ability to cap with dangerous force creating flying debris. process estradiol via cytoplasmic estrogen receptors and the 1. Thaw the vial by gentle agitation in a 37°C water bath. Growth of MCF7 cells is inhibited by tumor necrosis factor To reduce the possibility of contamination, keep the O- alpha (TNF alpha). Secretion of IGFBP's can be modulated ring and cap out of the water. Thawing should be rapid Karyology: modal number = 82; range = 66 to 87. The
stemline chromosome numbers ranged from hypertriploidy to 2. Remove the vial from the water bath as soon as the hypotetraploidy, with the 2S component occurring at 1%. contents are thawed, and decontaminate by dipping in or There were 29 to 34 marker chromosomes per S metaphase; spraying with 70% ethanol. All of the operations from this 24 to 28 markers occurred in at least 30% of cells, and point on should be carried out under strict aseptic generally one large submetacentric (M1) and 3 large subtelocentric (M2, M3, and M4) markers were recognizable 3. It is recommended that the cryoprotective agent be removed immediately. Centrifuge the cell suspension at
American Type Culture Collection
Product Information Sheet for ATCC® HTB-22™
approximately 125 xg for 5 to 10 minutes. Discard the Subculturing Procedure
supernatant and resuspend the cell pellet in an Volumes used in this protocol are for 75 cm2 flask; appropriate amount of fresh growth medium. proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. 4. Transfer the cell pellet to an appropriate size vessel. It Note: if floating cells are present, it is recommended that
is important to avoid excessive alkalinity of the medium they be transferred at the first two (2) subcultures as during recovery of the cells. It is suggested that, prior to described below. It is not necessary to transfer floating cells the addition of the vial contents, the culture vessel containing the growth medium be placed into the incubator for at least 15 minutes to allow the medium to 1. Remove culture medium to a centrifuge tube. reach its normal pH (7.0 to 7.6). 2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin - 0.53 5. Incubate the culture at 37°C in a suitable incubator. A mM EDTA solution to remove all traces of serum, which 5% CO2 in air atmosphere is recommended if using the medium described on this product sheet. 3. Add 2.0 to 3.0 ml of Trypsin-EDTA solution to flask and Note: Present batches of MCF7 cells are exhibiting the
observe cells under an inverted microscope until cell layer is dispersed (usually with 5 to 10 minutes). The cells usually attach as three-dimensional clusters and eventually grow to a 80-90% confluent monolayer. However, Note: To avoid clumping do not agitate the cells by
we are finding that most of the clusters remain in suspension hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed After first subculture all the cells will not attach. There will be clusters in suspension. Break up the clusters the best you can by gently pipetting with a small bore pipette (5 ml or 4. Add 6.0 to 8.0 ml of complete growth medium and smaller). After a few days incubation, the cells should reattach as three-dimensional islands (there will be some clusters that do not reattach). Growth will eventually spread 5. Transfer the cell suspension to the centrifuge tube with out from the islands and the culture should, after the second the medium and cells from Step #1 and spin at subculture, flatten and become 70-80% confluent. approximately 125 xg for 5 to10 minutes. Discard the Handling Procedure for Flask Cultures
The flask was seeded with cells (see specific batch 6. Resuspend the cell pellet in fresh growth medium. Add information) grown and completely filled with medium at appropriate aliquots of cell suspension to new culture ATCC to prevent loss of cells during shipping. vessels. Maintain cultures at a cell concentration between 2x10(4) and 2 x 10(5) cells/cm2. 1. Upon receipt visually examine the culture for Subcultivation Ratio: 1:3 to 1:6.
macroscopic evidence of any microbial contamination. Using an inverted microscope (preferably equipped with 7. Place culture vessels in incubators at 37°C. phase-contrast optics), carefully check for any evidence of microbial contamination. Also check to determine if Note: For more information on enzymatic dissociation
the majority of cells are still attached to the bottom of the and subculturing of cell lines consult Chapter 13 in Culture
flask; during shipping the cultures are sometimes Of Animal Cells: A Manual Of Basic Technique by R. Ian
handled roughly and many of the cells often detach and Freshney, 5th edition, published by Wiley-Liss, N.Y., 2005. become suspended in the culture medium (but are still Medium Renewal
2. If the cells are still attached, aseptically remove all but
5 to 10 ml of the shipping medium. The shipping medium Complete Growth Medium
can be saved for reuse. Incubate the cells at 37°C in a The base medium for this cell line is ATCC-formulated 5% CO2 in air atmosphere until they are ready to be Eagle's Minimum Essential Medium, Catalog No. 30-2003. To make the complete growth medium, add the following 3. If the cells are not attached, aseptically remove the
entire contents of the flask and centrifuge at 125 x g for • fetal bovine serum to a final concentration of 10% 5 to 10 minutes. Remove shipping medium and save. This medium is formulated for use with a 5% CO 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.
American Type Culture Collection
Product Information Sheet for ATCC® HTB-22™
ATCC tested fetal bovine serum is available as ATCC® is cell cycle regulated and determined by the nucleus.
Catalog No. 30-2020 (500ml) or ATCC® Catalog No. 30-2021 Cancer Res. 57: 5217-5220, 1997 PubMed: 98053886 van Dijk MA et al. A functional assay in yeas for
the human estrogen receptor displays wild-type and
Cryoprotectant Medium
variant estrogen receptor messenger RNAs present in
Complete growth medium described above supplemented breast carcinoma. Cancer Res. 57: 3478-3485, 1997
Cell culture tested DMSO is available as ATCC® Catalog No. Landers JE et al. Translational enhancement of
mdm2 oncogene expression in human tumor cells
containing a stabilized wild-type p53 protein. Cancer Res.
Additional Information
Umekita Y et al. Human prostate tumor growth in
Additional product and technical information can be obtained athymic mice: inhibition by androgens and stimulation
from the catalog references and the ATCC Web site at by finasteride. Proc. Natl. Acad. Sci. USA 93: 11802-11807,
Zamora-Leon SP et al. Expression of the fructose
References
transporter GLUT5 in human breast cancer. Proc. Natl.
(additional references may be available in the catalog at
Acad. Sci. USA 93: 1847-1852, 1996 PubMed: 96312501 Geiger T et al. Antitumor activity of a PKC-alpha
Sugarman BJ et al. Recombinant human tumor
antisense oligonucleotide in combination with standard
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chemotherapeutic agents against various human tumors
and transformed cells in vitro. Science 230: 943-945, 1985
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Takahashi K and Suzuki K. Association of insulin-
Jang SI et al. Activator protein 1 activity is
like growth-factor-I-induced DNA synthesis with
involved in the regulation of the cell type-specific
phosphorylation and nuclear exclusion of p53 in human
expression from the proximal promoter of the human
breast cancer MCF-7 cells. Int. J. Cancer 55: 453-458, 1993
profilaggrin gene. J. Biol. Chem. 271: 24105-24114, 1996
Brandes LJ and Hermonat MW. Receptor status
Lee JH et al. The proximal promoter of the
and subsequent sensitivity of subclones of MCF-7
human transglutaminase 3 gene. J. Biol. Chem. 271: 4561-
human breast cancer cells surviving exposure to
diethylstilbestrol. Cancer Res. 43: 2831-2835, 1983
Chang K and Pastan I. Molecular cloning of
mesothelin, a differentiation antigen present on
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Bacus SS et al. Differentiation of cultured human
accumulation of insulin-like growth factor-binding
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Soule HD et al. A human cell line from a pleural
Tissue Culture Methods 11:223-227, 1988. effusion derived from a breast carcinoma. J. Natl. Cancer
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ATCC Warranty
transcribed in trophoblastic cells. Cancer Res. 57: 516-
The viability of ATCC products is warranted for 30 days from the date of shipment. If you feel there is a problem with this Littlewood-Evans AJ et al. The osteoclast-
product, contact Technical Services by phone at 800-638- associated protease cathepsin K is expressed in human
6597 or 703-365-2700 or by e-mail at tech@atcc.org. Or you breast carcinoma. Cancer Res. 57: 5386-5390, 1997
Komarova EA et al. Intracellular localization of
p53 tumor suppressor protein in gamma-irradiated cells

American Type Culture Collection
Product Information Sheet for ATCC® HTB-22™
Disclaimers
This product is intended for laboratory research purposes
only. It is not intended for use in humans.
While ATCC uses reasonable efforts to include accurate and
up-to-date information on this product sheet, ATCC makes
no warranties or representations as to its accuracy. Citations
from scientific literature and patents are provided for
informational purposes only. ATCC does not warrant that
such information has been confirmed to be accurate.
This product is sent with the condition that you are
responsible for its safe storage, handling, and use. ATCC is
not liable for any damages or injuries arising from receipt
and/or use of this product. While reasonable effort is made to
insure authenticity and reliability of strains on deposit, ATCC
is not liable for damages arising from the misidentification or
misrepresentation of cultures.
Please see the enclosed Material Transfer Agreement (MTA)
for further details regarding the use of this product. The
MTA is also available on our W.
ATCC 2009. All rights reserved.
ATCC® is a registered trademark of the American Type
Culture Collection.
05/09


American Type Culture Collection

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