UWB1.289+BRCA1 (ATCC® CRL-2946)

Organism: Homo sapiens, human  /  Tissue: ovary  /  Disease: ovarian carcinoma

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Organism Homo sapiens, human
Tissue ovary
Product Format frozen
Morphology epithelial-like
Culture Properties adherent
Biosafety Level 2
Disease ovarian carcinoma
Age 56
Gender female
Storage Conditions liquid nitrogen vapor phase
Images
Derivation
UWB1.289+BRCA1 is a stable cell line derived from UWB1.289 (CRL-2945), a BRCA1-null human ovarian cancer line, in which wild-type BRCA1 was restored.
Clinical Data
female
Antigen Expression
BRCA1, positive
cytokeratin 7 (CK-7), positive
calretinin, positive
Wilms' tumor protein (WT), positive
Receptor Expression
estrogen, not expressed
progesterone, not expressed
Oncogene p53
Genes Expressed
p53, cytokeratin 7 (CK-7)
Comments

A pcDNA3 plasmid carrying wild-type BRCA1 was transfected into the parent line. Restoration of wild-type BRCA1 function in these cells partially restores DNA damage responses. RefDelloRusso, C., et al. Functional characterization of a novel BRCA1-null ovarian cancer cell line in response to ionizing radiation. Mol Cancer Res.;5(1):35-45, 2007. PubMed 17259345

Restoration of wild-type BRCA1 function in these cells partially restores DNA damage responses.

Complete Growth Medium The base medium for this cell line is:
  • 50% ATCC-formulated RPMI-1640 Medium, Catalog No. 30-2001.
  • 50% MEGM (Mammary Epithelial Growth Medium from Clonetics/Lonza (MEGM Bullet Kit; CC-3150) made of MEBM basal medium and SingleQuot additives (ATCC does not use gentamycin-amphotericin B). Note: Do not filter complete medium. To make the final complete growth medium add the following components to the base medium:
  • G-418 to a final concentration of 200ug/ml.
  • fetal bovine serum to a final concentration of 3%.
  • 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 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 2.0 to 3.0 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 culture vessels. An inoculum of 5 x 103 to 7 X 103 viable cells/cm2 is recommended.
    7. Incubate cultures at 37°C. Subculture when cell concentration is between 4 x 104 and 6 x 104 cells/cm2.

    Subcultivation ratio: A subcultivation ratio of 1:4 to 1:6 is recommended.
    Medium renewal: Every 2 to 3 days

    Cryopreservation
    Freeze medium: complete growth medium, 95%; DMSO, 5%
    Storage temperature: liquid nitrogen vapor phase
    Culture Conditions
    Atmosphere: 5% CO2 in air recommended
    Temperature: 37°C
    STR Profile
    D5S818: 13
    D13S317: 9
    D7S820: 7, 10
    D16S539: 12
    vWA: 16, 19
    THO1: 9
    TPOX: 9, 11
    CSF1PO: 11
    Amelogenin: X
    Population Doubling Time approximately 36 hours
    Name of Depositor E Swisher
    Year of Origin 2003
    References

    DelloRusso, C., et al. Functional characterization of a novel BRCA1-null ovarian cancer cell line in response to ionizing radiation. Mol Cancer Res.;5(1):35-45, 2007. PubMed 17259345

    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

    DelloRusso, C., et al. Functional characterization of a novel BRCA1-null ovarian cancer cell line in response to ionizing radiation. Mol Cancer Res.;5(1):35-45, 2007. PubMed 17259345