HCC1008 (ATCC® CRL-2320)

Organism: Homo sapiens, human  /  Cell Type: Epithelial  /  Tissue: breast; mammary gland/duct; derived from metastatic site: lymph node  /  Disease: TNM stage IIA, grade 3, ductal carcinoma

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Organism Homo sapiens, human
Tissue breast; mammary gland/duct; derived from metastatic site: lymph node
Cell Type Epithelial
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 1
Disease TNM stage IIA, grade 3, ductal carcinoma
Age 67 years adult
Gender female
Ethnicity Black
Storage Conditions liquid nitrogen vapor phase
Karyotype multiploid; cell population has several ploidy indices; double minute (DM) chromosomes were observed
Derivation

This cell line was initiated from an axillary lymph node on 6/7/94 and took 12.5 months to establish.

A B lymphoblastoid cell line initiated by Epstein-Barr virus (EBV) transformation of peripheral blood lymphocytes from this patient is available as HCC1007 BL (CRL-2319)

Clinical Data
67 years
female
Black
Receptor Expression
estrogen receptor, not expressed
progesterone receptor, not expressed
Oncogene her2/neu +, p53 +
Genes Expressed
Epithelial glycoprotein 2 (EGP2)
cytokeratin 19
Cellular Products
Epithelial glycoprotein 2 (EGP2)
cytokeratin 19
Comments

The cells are positive for expressions of Her2-neu and p53 oncogenes.

HCC1008 is positive for the epithelial cell specific marker, Epithelial Glycoprotein 2 (EGP2) and cytokeratin 19.

The cells are negative for expression of estrogen receptor (ER) and for expression of progesterone receptor (PR).

A B lymphoblastoid cell line initiated by Epstein-Barr virus (EBV) transformation of peripheral blood lymphocytes from this patient is available as HCC1007 BL (ATCC CRL-2319)

Complete Growth Medium HITES medium supplemented with 10% fetal bovine serum.The base medium for this cell line is ATCC-formulated DMEM:F12 Medium Catalog No.30-2006. To make the complete growth medium,add the following components to the base medium:
  • 0.005 mg/ml Insulin
  • 0.01 mg/ml Transferrin
  • 30nM Sodium selenite (final conc.)
  • 10 nM Hydrocortisone (final conc.)
  • 10 nM beta-estradiol (final conc.)
  • extra 2mM L-glutamine (for final conc. of 4.5 mM)
  • 10% fetal bovine serum (final conc.)

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. Add 3.0 to 4.0 mL Cell Dissociation Buffer (GIBCO #13150-016) to cell layer and incubate at room temperature or 37C.
  3. Observe cells under an inverted microscope until cell layer is dispersed (usually with 5 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 Cell Dissociation Buffer, transfer cell suspension to centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes.
  6. Discard supernatant and resuspend cell pellet in fresh complete growth medium. Aspirate cells with a small bore pipette. Add appropriate aliquots of cell suspension to new culture vessels.
  7. Place culture vessels in incubators at 37°C.

Note: Cells reattach slowly after subculture (about 50% attached and 50% in suspension). Allow flasks to remain undisturbed for at least a week for cultures to become reestablished. It can take as along as three to four weeks before cultures can be subcultured again.

Subcultivation Ratio: A subcultivation ratio of 1:2 is recommended
Medium Renewal: Every 3 to 4 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 7.5% (v/v) DMSO. Cell culture tested DMSO is available as ATCC Catalog No. 4-X.
Culture Conditions
Temperature: 37°C
Atmosphere: Air, 95%; Carbon dioxide (CO2), 5% DMSO
Note: This cell line grows exceedingly slow. Cells grow in patches and cultures only become 50 to 60% confluent.
STR Profile
Amelogenin: X
CSF1PO: 12,13
D13S317: 12
D16S539: 14,15
D5S818: 13
D7S820: 10
THO1: 7
TPOX: 11
vWA: 17,19
Name of Depositor AF Gazdar, AK Virmani
Year of Origin June 7, 1994
References

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

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.

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
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

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

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.