UWB1.289+BRCA1細胞

數(shù)量： 大量

器官來源： 卵巢

是否是腫瘤細胞： 0

物種來源： 人

ATCC Number： CRL-2946?

相關**： 其他**

細胞形態(tài)： 上皮樣

運輸方式： 凍存運輸

生長狀態(tài)： 貼壁生長

年限： 56

UWB1.289+BRCA1細胞Designations: UWB1.289+BRCA1

Depositors: E. Swisher

Biosafety Level: 2

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: BRCA1, positive

cytokeratin 7 (CK-7), positive

calretinin, positive

Wilms' tumor protein (WT), positive

DNA Profile (STR): D5S818: 13

D13S317: 9

D7S820: 7, 10

D16S539: 12

vWA: 16, 19

THO1: 9

UWB1.289+BRCA1細胞TPOX: 9, 11

CSF1PO: 11

Amelogenin: X

Age: 56

Gender: female

Comments: 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. 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. 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 (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%.

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: UWB1.289+BRCA1細胞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 growth medium, 95%; DMSO, 5%

Storage temperature: liquid nitrogen vapor phase

Doubling Time: approximately 36 hours

Related Products: parental cell line - ATCC CRL-2945 (UWB1.2

Recommended medium: 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: UWB1.289+BRCA1細胞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