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
commercial use of any cell line first contact the originating
investigator to negotiate an agreement. Third party
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
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
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°Cin 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 cycleregulated 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 necrosis factor-alpha: effects on proliferation of normal chemotherapeutic agents against various human tumors and transformed cells in vitro. Science 230: 943-945, 1985 transplanted into nude mice. Anti-Cancer Drug Des. 13:
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
Lan MS et al. Polypeptide core of a human mesothelium, mesotheliomas, and ovarian cancers. Proc. pancreatic tumor mucin antigen. Cancer Res. 50: 2997-
Natl. Acad. Sci. USA 93: 136-140, 1996 PubMed: 96133892
Zhu X et al. Cell cycle-dependent modulation of
Pratt SE and Pollak MN. Estrogen and telomerase activity in tumor cells. Proc. Natl. Acad. Sci. antiestrogen modulation of MCF7 human breast cancer
USA 93: 6091-6095, 1996 PubMed: 96234095
cell proliferation is associated with specific alterations in
Bacus SS et al. Differentiation of cultured human accumulation of insulin-like growth factor-binding breast cancer cells (AU-565 and MCF-7) associated with proteins in conditioned media. Cancer Res. 53: 5193- loss of cell surface HER-2/neu antigen. Mol. Carcinog. 3:
Huguet EL et al. Differential expression of human
Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992),
Wnt genes 2, 3, 4, and 7B in human breast cell lines and ATCC Quality Control Methods for Cell Lines. 2nd edition, normal and disease states of human breast tissue.
Cancer Res. 54: 2615-2621, 1994 PubMed: 94221588
Caputo, J. L., Biosafety procedures in cell culture. J.
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
Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley,
Inst. 51: 1409-1416, 1973 PubMed: 74054239
D., (1995) Laboratory Safety: Principles and Practice.
Bellet D et al. Malignant transformation of
Second edition, ASM press, Washington, DC.
nontrophoblastic cells is associated with the expression of chorionic gonadotropin beta genes normally 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 email@example.com. 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
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