HCC1937 (ATCC® CRL-2336)

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

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Organism Homo sapiens, human
Tissue mammary gland; breast/duct
Cell Type lymphoblast, epithelial
Product Format frozen
Morphology epithelial
Culture Properties adherent, The line grows as large epithelial cells with a tendency to float at high cell densities
Biosafety Level 1
Disease TNM stage IIB, grade 3,primary ductal carcinoma
Age 23 years adult
Gender female
Ethnicity Caucasian
Storage Conditions liquid nitrogen vapor phase
Karyotype CRL-2336 is highly transformed which is evident from the chromosome count and karyotype description. The modal chromosome number is 100. At least forty-three marker chromosomes, involving nearly every chromosome, were found. Chromosome 1 and chromosome 3 derivative chromosomes were verified using commercial whole chromosome paint (fluorescent in-situ hybridization (FISH)) probes. An acrocentric chromosome with an extra C-band at qter was detected (2 copies per metaphase). There were no normal X chromosomes however at least one derivative X was seen in each cell. The absence of a Y chromosome was verified by QM staining and C-bands. Normal copies of N1, N4, N15, N16 and N18 were absent. More detailed cytogenetic information is available upon request.
Derivation
This cell line was initiated from a primary ductal carcinoma on October 13, 1995, and took 11.5 months to establish

Clinical Data
23 years adult
Caucasian
female
Receptor Expression
estrogen receptor, negative
progesterone receptor, negative
Oncogene BRCA1 (mutated, insertion C at nucleotide 5382), her2/neu -, p53 -
Genes Expressed
Epithelial glycoprotein 2 (EGP2)
cytokeratin 19
Comments
The tumor was classified as TNM Stage IIB, grade 3

BRCA1 analysis revealed that the cell line is homozygous for the BRCA1 5382C mutation, whereas the lymphoblastoid cell line derived from the same patient is heterozygous for the same mutation

This mutation was present in two other family members; an identical sister also developed breast cancer

The cell line has an acquired mutation of TP53 with wild-type allele loss; an acquired homozygous deletion of the PTEN gene, and loss of heterozygosity at multiple loci known to be involved in the pathogenesis of breast cancer

The cells are negative for expression of Her2-neu and for expression of p53.
HCC1937 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 progesterone receptor (PR)

An EBV transformed lymphoblastoid cell line (HCC1937BL) from the same patient is available as ATCC CRL-2337


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: 1:2 to 1:4
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
STR Profile
Amelogenin: X
CSF1PO: 12
D13S317: 13
D16S539: 13,14
D5S818: 12
D7S820: 9,10
THO1: 6
TPOX: 11
vWA: 16,17
Name of Depositor AF Gazdar, AK Virmani
Year of Origin October 13, 1995
References

Tomlinson GE, et al. Characterization of a breast cancer cell line derived from a germ-line BRCA1 mutation. Cancer Res. 58: 3237-3242, 1998. PubMed: 9699648

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
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

Tomlinson GE, et al. Characterization of a breast cancer cell line derived from a germ-line BRCA1 mutation. Cancer Res. 58: 3237-3242, 1998. PubMed: 9699648

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