UACC-3199 (ATCC® CRL-2983)

Organism: Homo sapiens, human  /  Tissue: mammary gland/breast duct; derived from metastatic site: axillary lymph node  /  Disease: infiltrating ductal carcinoma of the breast

Permits and Restrictions

View Permits

Organism Homo sapiens, human
Tissue mammary gland/breast duct; derived from metastatic site: axillary lymph node
Product Format frozen
Morphology epithelial-like
Culture Properties adherent
Biosafety Level 1
Disease infiltrating ductal carcinoma of the breast
Age 58 years old
Gender female
Ethnicity Caucasian
Applications
This cell line is an excellent tool for research into aberrant cytosine methylation of CpG island sites and BRCA1 repression in sporadic breast cancer. It is also very useful as a methylated BRCA1 control.
Storage Conditions liquid nitrogen vapor phase
Images
Derivation
UACC-3199 was derived from a 58 year-old female with infiltrating ductal carcinoma of the breast metastatic to the left axillary lymph nodes. 
Clinical Data
female
The patient had a family history of breast cancer.
58 years old
Caucasian
Receptor Expression
epidermal growth factor receptor (EGF), expressed
estrogen receptor, not expressed
progesterone receptor, not expressed
Oncogene c-erbB-2 positive
Genes Expressed
c-erbB-2 positive
Tumorigenic YES
Comments The patient had a family history of breast cancer.
Complete Growth Medium The base medium for this cell line is ATCC-formulated Leibovitz's L-15 Medium, Catalog No. 30-2008. To make the complete growth medium, add the following components to the base medium:
  • fetal bovine serum to a final concentration of 5%
  • 0.01 mg/ml transferrin (final conc.)
  • 0.01 mg/ml insulin (final conc.)
  • 5 µg/ml (55 U/ml) catalase (final conc.)
  • 3.6 µg/ml (0.01 mM) hydrocortisone (final conc.)
  • extra 2 mM glutamine

Subculturing
Volumes used in this protocol are for 75 cm2 flasks; 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 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 1.0 to 2.0 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. Transfer cell suspension to a centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes. Discard supernatant.
  6. Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new culture vessels. An inoculum of 1 X 104 to 2 X 104 viable cells/cm2 is recommended.
  7. Incubate cultures at 37°C. Subculture when the cell concentration is between 2 X 104 to 5 X 104 cells/cm2.
Subcultivation ratio: A subcultivation ratio of 1:2 to 1:3 is recommended
Medium renewal: Every 3 to 4 days
Cryopreservation
Freeze medium: complete growth medium, 90%; DMSO, 10%
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Temperature: 37°C
Atmosphere: air, 100%
Growth Conditions: Cells grow very slowly. Cultures reach confluence in 2 to 3 weeks.
Growth Conditions: Avoid inoculating vessels below recommended seeding density.
STR Profile
CSF1PO: 12
D13S317: 11
D16S539: 12
D5S818: 13
D7S820: 10
THO1: 9.3
TPOX: 10, 13
vWA: 15, 17
Amelogenin: X
Name of Depositor K Brown
Year of Origin November 1994
References

Rice JC, et al. Aberrant methylation of the BRCA1 CpG island promoter is associated with decreased BRCA1 mRNA in sporadic breast cancer cells. Oncogene 17(14): 1807-1812, 1998. PubMed: 9778046

Rice JC, et al. Transcriptional repression of BRCA1 by aberrant cytosine methylation, histone hypoacetylation and chromatin condensation of the BRCA1 promoter. Nucleic Acids Res. 28(17): 3233-3239, 2000. PubMed: 10954590

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
References

Rice JC, et al. Aberrant methylation of the BRCA1 CpG island promoter is associated with decreased BRCA1 mRNA in sporadic breast cancer cells. Oncogene 17(14): 1807-1812, 1998. PubMed: 9778046

Rice JC, et al. Transcriptional repression of BRCA1 by aberrant cytosine methylation, histone hypoacetylation and chromatin condensation of the BRCA1 promoter. Nucleic Acids Res. 28(17): 3233-3239, 2000. PubMed: 10954590