產(chǎn)品名稱 |
283TAg |
商品貨號 |
B206505 |
Organism |
Mus musculus, mouse |
Tissue |
embryo |
Cell Type |
fibroblast immortalized with SV40 large T antigenSV40 large T antigen transfected |
Product Format |
frozen |
Morphology |
fibroblast |
Culture Properties |
adherent |
Biosafety Level |
2 cells containing SV40 viral 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. |
Age |
14.5 day gestation embryo |
Applications |
DNA repair studies |
Storage Conditions |
liquid nitrogen vapor phase |
Images |
 |
Derivation |
283TAg (Polb/Mpg double null) is a mouse embryonic fibroblast (MEF) cell line derived from polymerase (DNA directed), beta (Polb)/ N-methylpurine-DNA glycosylase (Mpg, Aag) double null embryos at day 14.5 of gestation.
The cell line was established by transfection with an expression vector for SV40 large T antigen [PubMed: 8538772]. The cells are transgenic for lambda LIZ (Lac I/cII). |
Comments |
283TAg (Polb/Mpg double null) is a mouse embryonic fibroblast (MEF) cell line derived from polymerase (DNA directed), beta (Polb)/ N-methylpurine-DNA glycosylase (Mpg, Aag) double null embryos at day 14.5 of gestation.
The cell line was established by transfection with an expression vector for SV40 large T antigen [PubMed: 8538772]. The cells are transgenic for lambda LIZ (Lac I/cII). |
Complete Growth Medium |
The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
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Subculturing |
Protocol: - Remove and discard culture medium.
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Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
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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 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
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Add appropriate aliquots of the cell suspension to new culture vessels.
An inoculum of 4 X 10(3) to 4 X 10(4) viable cells/cm2 is recommended. - Incubate cultures at 37°C.
Interval: Maintain cultures at a cell concentration between 6 X 10(3) and 1 X 10(5) cells/cm2. Subcultivation Ratio: A subcultivation ratio of 1:6 to 1:8 is recommended Medium Renewal: Two to three times weekly |
Cryopreservation |
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase |
Culture Conditions |
Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37.0°C |
Population Doubling Time |
29 hours |
Name of Depositor |
RW Sobol |
Year of Origin |
2000 |
References |
Sobol RW, et al. Requirement of mammalian DNA polymerase-beta in base-excision repair. Nature 379: 183-186, 1996. PubMed: 8538772
Sobol RW, et al. Base excision repair intermediates induce p53-independent cytotoxic and genotoxic responses. J. Biol. Chem. 278: 39951-39959, 2003. PubMed: 12882965
Engelward BP, et al. Base excision repair deficient mice lacking the Aag alkyladenine DNA glycosylase. Proc. Natl. Acad. Sci. USA 94: 13087-13092, 1997. PubMed: 9371804
Sobol RW, et al. Mutations associated with base excision repair deficiency and methylation-induced genotoxic stress. Proc. Natl. Acad. Sci. USA 99: 6860-6865, 2002. PubMed: 11983862
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