HCC1428 (ATCC® CRL-2327)

Organism: Homo sapiens, human  /  Cell Type: Epithelial  /  Tissue: mammary gland/breast; derived from metastatic site: adenocarcinoma and pleural effusion  /  Disease: TNM stage IV, grade 4, adenocarcinoma

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Organism Homo sapiens, human
Tissue mammary gland/breast; derived from metastatic site: adenocarcinoma and pleural effusion
Cell Type Epithelial
Product Format frozen
Morphology epithelial
Culture Properties adherent; large epithelial cells with occasional vacuole formation
Biosafety Level 1
Disease TNM stage IV, grade 4, adenocarcinoma
Age 49 years
Gender female
Ethnicity Caucasian, White
Storage Conditions liquid nitrogen vapor phase
Karyotype polyploid
Derivation
HCC1428 was derived from a female who harbored a homozygous deletion in the FHIT gene.
This cell line was initiated on 11/4/95 and took 15 months to establish.
Clinical Data
female
Caucasian, White
49 years
There is a family history of breast cancer (maternal grandmother).
Oncogene her2/neu -, p53 -
Genes Expressed
HCC1428 cells are positive for the expression of Epithelial glycoprotein 2 [EGP2]; cytokeratin 19
The cells are negative for expression of Her2-neu and for expression of p53.
Comments

An EBV transformed lymphoblastoid cell line [HCC1428 BL] from the same patient is available as ATCC CRL-2328. The homozygous deletion in the FHIT gene is retained in the lymphoblastoid line.

The FHIT gene, which spans the FRA3B fragile site at chromosome 3p14.2, is a candidate tumor suppressor gene in breast and other cancers.

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 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 1:8
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 95%; DMSO, 5%
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Temperature: 37°C
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
STR Profile
CSF1PO: 10
D13S317: 12
D16S539: 12,8
D5S818: 11,12
D7S820: 9
THO1: 8
TPOX: 8
vWA: 17
Name of Depositor AF Gazdar, AK Virmani
Year of Origin November 4, 1995
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

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