UACC-1179 (ATCC® CRL-3127)

Organism: Homo sapiens, human  /  Cell Type: epithelial-like  /  Tissue: breast
Derived from metastatic site: pleural fluid
 /  Disease: adenocarcinoma

Organism Homo sapiens, human
Tissue breast
Derived from metastatic site: pleural fluid
Cell Type epithelial-like
Product Format frozen
Morphology epithelial-like
Culture Properties adherent
Biosafety Level 1
Disease adenocarcinoma
Age 61 years old
Gender female
Applications
Theses cells have been used in studies of the p53 mutation, as well as the regulation of p53 on DSC3 and MAPSIN.
Storage Conditions liquid nitrogen vapor phase
Karyotype range=32-75; modal number=34-35; 9pHSR
Images
Derivation UACC-1179 was derived from a 61 year-old female with breast cancer. The patient underwent chemotherapy
Receptor Expression
neu (ErbB2; HER2), expressed
epidermal growth factor receptor (EGFR), not expressed
estrogen receptor (ER), not expressed
progesterone receptor (PR), not expressed
Comments
UACC-1179 is negative for tumor suppressor genes MASPIN and the cell adhesion molecule DSC3.
Inhibition of DNA methyltransferase could induce transcriptional reactivation of both DSC3 and MASPIN of this cell line.

The p53R213X mutation has been reported in this cell line. 


The cells have been used in studies of p53 mutation, as well as the relegation of p53 on DSC3 and MAPSIN.
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
These cells are cultured on collagen I coated vessels. Add 5 µg per cm2 collagen I (BD Biosciences, Cat. No.354236 or equivalent) to culture vessels and incubate at room temperature for 1 hour. Remove collagen solution and rinse vessels 3 times with a balanced salt solution. Vessels may be used immediately or air dried and stored at 2-8°C for up to one week under sterile conditions. Alternatively, commercially available pre-coated Collagen I vessels, such as BD BioCoat Cellware (BD Biosciences, Cat. No. 356524 for 75 cm2 flask) or equivalent, may be used.
Volumes used in this protocol are for 75 cm2 flasks; proportionally reduce or increase amount of solutions 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 (DPBS) 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 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 collagen I coated culture vessels. An inoculum of 3 x 104 to 4 x 104 viable cells/cm2 is recommended.
  7. Incubate cultures at 37°C.
Subculture when cultures reach a cell concentration between 7 x 104 to 1 x 105cells/cm2
Subcultivation ratio: A subcultivation ratio of 1:2 is recommended.
Medium renewal: every 2 to 3 days
Cryopreservation
Freeze medium: complete growth medium, 80%; FBS, 10%; DMSO, 10%
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Temperature: 37.0°C
Atmosphere: air, 100%
Growth Conditions: This cell line grows slowly.
STR Profile
D5S818: 12
D13S317: 12
D7S820: 8, 11
D16S539: 10, 13
vWA: 16
TH01: 6
Amelogenin: X
TPOX: 8
CSF1PO: 10
Name of Depositor K Brown
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

Domann FE, et al. Epigenetic silencing of maspin gene expression in human breast cancers. Int. J. Cancer. 85(6): 805-810, 2000 PubMed: 10709100

Oshiro MM, et al. Mutant p53 and aberrant cytosine methylation cooperate to silence gene expression. Oncogene 22(23): 3624-3634, 2003. PubMed: 12789271

Watts GS, et al. The acetyltransferase p300/CBP-associated factor is a p53 target gene in breast tumor cells. Neoplasia 6(3): 187-194, 2004. PubMed: 15153330

Futscher BW, et al. Chapter 6: Epigenetic Dysregulation of Maspin (SerpinB5) in Cancer Invasion and Metastasis. DNA Methylation, Epigenetics and Metastasis. Cancer Metastasis - Biology and Treatment, 7: 133-155, 2005.

Oshiro MM, et al. Epigenetic silencing of DSC3 is a common event in human breast cancer. Breast Cancer Res. 7(5): R669-R680, 2005. PubMed: 16168112

Futscher BW, et al. Chapter 7: Epigenetic Regulation of Genes That Affect Tumor Cell Adhesion. Cell Adhesion and Cytoskeletal Molecules in Metastasis. Cancer Metastasis - Biology and Treatment, 9: 123-140, 2006.

Wozniak RJ, et al. 5-Aza-2'-deoxycytidine-mediated reductions in G9A histone methyltransferase and histone H3 K9 di-methylation levels are linked to tumor suppressor gene reactivation. Oncogene 26(1): 77-90, 2007. PubMed: 16799634

Robey IF, et al. Regulation of the warburg effect in early-passage breast cancer cells. Neoplasia 10(8): 745-756, 2008. PubMed: 18670636

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

Domann FE, et al. Epigenetic silencing of maspin gene expression in human breast cancers. Int. J. Cancer. 85(6): 805-810, 2000 PubMed: 10709100

Oshiro MM, et al. Mutant p53 and aberrant cytosine methylation cooperate to silence gene expression. Oncogene 22(23): 3624-3634, 2003. PubMed: 12789271

Watts GS, et al. The acetyltransferase p300/CBP-associated factor is a p53 target gene in breast tumor cells. Neoplasia 6(3): 187-194, 2004. PubMed: 15153330

Futscher BW, et al. Chapter 6: Epigenetic Dysregulation of Maspin (SerpinB5) in Cancer Invasion and Metastasis. DNA Methylation, Epigenetics and Metastasis. Cancer Metastasis - Biology and Treatment, 7: 133-155, 2005.

Oshiro MM, et al. Epigenetic silencing of DSC3 is a common event in human breast cancer. Breast Cancer Res. 7(5): R669-R680, 2005. PubMed: 16168112

Futscher BW, et al. Chapter 7: Epigenetic Regulation of Genes That Affect Tumor Cell Adhesion. Cell Adhesion and Cytoskeletal Molecules in Metastasis. Cancer Metastasis - Biology and Treatment, 9: 123-140, 2006.

Wozniak RJ, et al. 5-Aza-2'-deoxycytidine-mediated reductions in G9A histone methyltransferase and histone H3 K9 di-methylation levels are linked to tumor suppressor gene reactivation. Oncogene 26(1): 77-90, 2007. PubMed: 16799634

Robey IF, et al. Regulation of the warburg effect in early-passage breast cancer cells. Neoplasia 10(8): 745-756, 2008. PubMed: 18670636