產(chǎn)品名稱 |
UWB1.289+BRCA1 |
商品貨號 |
B224800 |
Organism |
Homo sapiens, human |
Tissue |
ovary |
Product Format |
frozen |
Morphology |
epithelial-like |
Culture Properties |
adherent |
Biosafety Level |
2 [Cells contain CMV and SV40 vial DNA sequences]
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. |
Disease |
ovarian carcinoma |
Age |
56 |
Gender |
female |
Storage Conditions |
liquid nitrogen vapor phase |
Images |
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Derivation |
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. |
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%.
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Subculturing |
Volumes used in this protocol are for 75 cm 2 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 37°C 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 103 to 7 X 103 viable cells/cm2 is recommended.
- 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
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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
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