HCC1569 (ATCC® CRL-2330)

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

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Organism Homo sapiens, human
Tissue mammary gland; breast
Cell Type Epithelial
Product Format frozen
Morphology epithelial
Culture Properties mixed adherent-suspension
Biosafety Level 1
Disease TNM stage IV, grade 3, primary metaplastic carcinoma
Age 70 years
Gender female
Ethnicity Black
Karyotype polyploid
Derivation
The cell line is derived from an older patient with a germline mutation in the FHIT gene. The FHIT gene is located at 3p14.2 and the mutation is a transversion at nucleotide 651 (G to T).The patient's daughter carries the same gene mutation.
This cell line was initiated on 3/8/95 and took 19 months to establish.
Clinical Data
70 years
Black
female
The patient received prior chemotherapy and had no family history of breast cancer.
The tumor was classified as TNM stage IV, grade 3, metaplastic carcinoma with 4 out of 18 lymph node metastasis.
Receptor Expression
estrogen receptor, negative
progesterone receptor, negative
Oncogene her2/neu +, p53 -
Genes Expressed
Epithelial glycoprotein 2 [EGP2]; cytokeratin 19
Cellular Products
Epithelial glycoprotein 2 [EGP2]; cytokeratin 19
Comments
The patient received prior chemotherapy and had no family history of breast cancer. The tumor was classified as TNM stage IV, grade 3, metaplastic carcinoma with 4 out of 18 lymph node metastasis.

The cells are poorly differentiated.

The cells are positive for expression of Her2-neu and negative for expression of p53.

HCC1569 is positive for the epithelial cell specific marker Epithelial Glycoprotein 2 (EGP2) and for cytokeratin 19.

The cells are negative for expression of estrogen receptor (ER) and for expression of progesterone receptor (PR) by immunohistochemistry, but PR expression was detectable at a low level by cytosolic protein assay.

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%.
Subculturing Volumes used in this protocol are for a 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.
  1. Remove culture medium, which contains suspended cells, to a centrifuge tube.
  2. Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.25% (w/v) Trypsin-0.53 mM 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 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.

  1. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting. 
  2. To remove trypsin-EDTA solution, transfer cell suspension to the centrifuge tube containing the medium and cells from step #1 and centrifuge at approximately 125 x g for 5 to 10 minutes.
  3. Discard supernatant and resuspend cells in fresh growth medium.  Add appropriate aliquots of cell suspension to new culture vessels.
  4. Incubate cultures at 37°C.

Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:6 is recommended
Medium Renewal: Every 2 to 3 days. 
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 13 in Culture Of Animal Cells: A Manual Of Basic Technique by R. Ian Freshney, 5th edition, published by Wiley-Liss, N.Y., 2005.
Cryopreservation
culture medium 95%; DMSO, 5%
Culture Conditions
Temperature: 37°C
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Name of Depositor AF Gazdar, AK Virmani
References

Ahmadian M, et al. Analysis of the FHIT gene and FRA3B region in sporadic breast cancer, preneoplastic lesions, and familial breast cancer probands. Cancer Res. 57: 3664-3668, 1997. PubMed: 9288768

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
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

Ahmadian M, et al. Analysis of the FHIT gene and FRA3B region in sporadic breast cancer, preneoplastic lesions, and familial breast cancer probands. Cancer Res. 57: 3664-3668, 1997. PubMed: 9288768

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