UWB1.289細胞

生長狀態(tài)： 貼壁生長

年限： 56

是否是腫瘤細胞： 0

物種來源： 人

ATCC Number： CRL-2945?

器官來源： 卵巢

相關**： 其他**

數量： 大量

運輸方式： 凍存運輸

細胞形態(tài)： 上皮樣

Designations: UWB1.289

Depositors: E. Swisher

UWB1.289細胞Biosafety Level: 1

Shipped: frozen

Medium & Serum: See Propagation

Growth Properties: adherent

Organism: Homo sapiens

Morphology: epithelial-like

Source: Organ: ovary

Disease: ovarian carcinoma

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Isolation: Isolation date: 2003

Receptors: estrogen, not expressed

progesterone, not expressed

Oncogene: p53

Antigen Expression: cytokeratin 7 (CK-7), positive

calretinin, positive

Wilms' tumor protein (WT), positive

BRCA1, negative

DNA Profile (STR): UWB1.289細胞D5S818: 13

D13S317: 9

D7S820: 7,10

D16S539: 12

vWA: 16,19

THO1: 9

CSF1PO: 11

Amelogenin: X

TPOX: 9,11

Age: 56

Gender: female

Comments: 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. [PubMed 17259345].

Propagation: ATCC complete growth medium: The base medium for this cell line is:

50% ATCC-formulated RPMI-1640 Medium, Catalog No. 30-2001.

50% MEGM UWB1.289細胞(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%.

Atmosphere: 5% CO2 in air recommended

Temperature: 37.0°C

Subculturing: Protocol: Volumes used in this protocol are for 75 sq cm flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.

Remove and discard culture medium.

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.

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 37C to facilitate dispersal.

Add 2.0 to 3.0 ml of complete growth medium and aspirate cells by gently pipetting.

Transfer cell suspension to a centrifuge tube and spin at approximately 125 X g for 5 to 10 minutes. Discard supernatant.

Resuspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to culture vessels. An inoculum of 5 X 10(3) to 7 X 10(3) viable cells/sq. cm is recommended.

Incubate cultures at 37C. Subculture when cell concentration is between 4 X 10(4) and 6 X 10(4) cells/sq. cm.

Subcultivation ratio: A subcultivation ratio of 1:4 to 1:6 is recommended.

Medium renewal: Every 2 to 3 days

Preservation: Freeze medium: complete culture medium, 95%; DMSO, 5%

Storage temperature: liquid nitrogen vapor phase

Doubling Time: approximately 53 hours

Related Products: UWB1.289+BRCA1: ATCC CRL-2946

Recommended medium: UWB1.289細胞ATCC 30-2001

Recommended serum: ATCC 30-2020

0.25% (w/v) Trypsin - 0.53mM EDTA in Hank's BSS (w/o Ca++, Mg ++): ATCC 30-2101

Phosphate-buffered saline: ATCC 30-2200

Cell culture tested DMSO: ATCC 4-X

Erythrosin B vital stain solution: ATCC 30-2404

References: 16172667: 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