Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

UWB1.289+BRCA1 is a stable cell line derived from UWB1.289 (ATCC CRL-2945), a BRCA1-null human ovarian cancer line, in which wild-type BRCA1 was restored.

female

BRCA1, positive

cytokeratin 7 (CK-7), positive

calretinin, positive

Wilms tumor protein (WT), positive

estrogen, not expressed

progesterone, not expressed

p53, cytokeratin 7 (CK-7)

Volumes used in this protocol are for 75xa0cm² 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 Dulbeccos 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.

Medium renewal: Every 2 to 3 days

Freeze medium: complete growth medium, 95%; DMSO, 5%

Storage temperature: liquid nitrogen vapor phase

Atmosphere: 5% CO₂ in air recommended

Temperature: 37°C

D5S818: 13

D13S317: 9

D7S820: 7, 10

D16S539: 12

vWA: 16, 19

THO1: 9

TPOX: 9, 11

CSF1PO: 11

Amelogenin: X