HCC70 (ATCC® CRL-2315)

Organism: Homo sapiens, human  /  Cell Type: Epithelial  /  Tissue: breast; mammary gland/duct  /  Disease: TNM stage IIIA, grade 3, primary ductal carcinoma

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Organism Homo sapiens, human
Tissue breast; mammary gland/duct
Cell Type Epithelial
Product Format frozen
Morphology epithelial
Culture Properties adherent, The line grows as attached medium-sized epithelial cells.
Biosafety Level 1
Disease TNM stage IIIA, grade 3, primary ductal carcinoma
Age 49 years
Gender female
Ethnicity Black
Storage Conditions liquid nitrogen vapor phase
Karyotype polyploid
Images
Derivation
The HCC70 cell line was initiated from a primary ductal carcinoma on June 3, 1992, and took 44 months to establish.
Clinical Data
49 years
Black
female
Receptor Expression
progesterone receptor, not expressed
Oncogene her2/neu -, p53 + (overexpressed)
Genes Expressed
Epithelial glycoprotein 2 [EGP2]; cytokeratin 19
Comments

The cells are poorly differentiated.

The tumor was classified as TNM Stage IIIA, grade 3, invasive ductal carcinoma with metastases in 4 out of 17 lymph nodes.

The cells overexpress p53, and are negative for the expression of Her2-neu oncogenes.

HCC70 is positive for the epithelial cell specific marker Epithelial Glycoprotein 2 (EGP2) and for cytokeratin 19.

The depositor of ATCC CRL-2315 initially reported the cells were positive for estrogen receptors. However, recent studies performed outside of ATCC have given conflicting results about the status of the estrogen receptor for these cells. An independent study at ATCC confirmed that ATCC CRL-2315, HCC70, is ER negative, PR negative and Her2 negative.

Complete Growth Medium The base medium for this cell line is ATCC-formulated RPMI-1640 Medium, Catalog No. 30-2001. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.


Phenol Red is a structural mimic for estrogen. Therefore the estrogenic activity of phenol red should be considered in any studies that utilize estrogen-responsive cells in culture (RefBerthois Y, et al. Phenol red in tissue culture media is a weak estrogen: implications concerning the study of estrogen-responsive cells in culture. Proc. Natl. Acad. Sci. USA 83: 2496–2500, 1986. PubMed: 3458212). The Phenol Red-free version of RPMI 1640, is ATCC catalog No. 30-2602.

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-0.53 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 (usually within 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. Add appropriate aliquots of the cell suspension to new culture vessels.
  6. Incubate cultures at 37°C
Subculture Ratio: 1:4 to 1:6
Medium Renewal: Every 2 to 3 days.
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.
Cryopreservation
Freeze Medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage Temperature: Liquid nitrogen vapor phase
Culture Conditions
Atmosphere: Air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
STR Profile
Amelogenin:X
CSF1PO:10,14
D13S317:12
D16S539:9,13
D5S818:12,13
D7S820:10,11
THO1:9
TPOX:10
vWA: 13,15
Name of Depositor AF Gazdar, AK Virmani
Year of Origin 1992
References

Kao J, et al. Molecular profiling of breast cancer cell lines defines relevant tumor models and provides a resource for cancer gene discovery. PlosONE 4 (7): e 6146, 2009

Kim MS, et al. Breast cancer diagnosis using a microfluidic multiplexed immunohistochemistry platform. PlosONE 5 (5): e10441, 2010

Gazdar AF, et al. Characterization of paired tumor and non-tumor cell lines established from patients with breast cancer. Int. J. Cancer 78: 766-774, 1998. PubMed: 9833771

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
Other Documentation
Restrictions

The line is available with the following restrictions: 1. This cell line was deposited at the ATCC by Dr. Adi F. Gazdar and is provided for research purposes only. Neither the cell line nor products derived from it may be sold or used for commercial purposes. Nor can the cells be distributed to third parties for purposes of sale, or producing for sale, cells or their products. The cells are provided as service to the research community. They are provided without warranty of merchantability or fitness for a particular purpose or any other warranty, expressed or implied. 2. Any proposed commercial use of the these cells, or their products must first be negotiated with the University of Texas Southwestern Medical Center at Dallas, 5323 Harry Hines Blvd., Dallas, Texas 75235. Telephone (214) 648-1888, Email TechnologyDevelopment@UTSouthwestern.edu, or Fax: (214) 951-0935.

References

Kao J, et al. Molecular profiling of breast cancer cell lines defines relevant tumor models and provides a resource for cancer gene discovery. PlosONE 4 (7): e 6146, 2009

Kim MS, et al. Breast cancer diagnosis using a microfluidic multiplexed immunohistochemistry platform. PlosONE 5 (5): e10441, 2010

Gazdar AF, et al. Characterization of paired tumor and non-tumor cell lines established from patients with breast cancer. Int. J. Cancer 78: 766-774, 1998. PubMed: 9833771