BRCA1-null human ovarian cancer cell line UWB1.289 is from a tumor of papillary serous histology, the most common form of ovarian carcinoma.

The patient developed breast cancer at age 42, ovarian cancer at age 54, and died at age 56.

UWB1.289 carries a germline BRCA1 mutation within exon 11 and has a deletion of the wild-type allele.

It is estrogen and progesterone receptor negative and has an acquired somatic mutation in p53. It is sensitive to ionizing radiation. 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

• 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:
• fetal bovine serum to a final concentration of 3%.

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.

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