127TAg細胞
生長狀態： 貼壁生長
運輸方式： 凍存運輸
細胞形態： 成纖維樣
細胞類型： 其他細胞類型
是否是腫瘤細胞： 0
物種來源： 小鼠
ATCC Number： CRL-2817?
數量： 大量
年限： 14.5 days gestation embryo
器官來源： 胚胎
Designations: 127TAg
127TAg細胞Depositors: RW Sobol
Biosafety Level: 2 [cells containing SV40 viral DNA sequences ]
Shipped: frozen
Medium & Serum: See Propagation
Growth Properties: adherent
Organism: Mus musculus
Morphology: fibroblast
Source: Organ: embryo
Cell Type: fibroblast immortalized with SV40 large T antigenSV40 large T antigen transfected
Permits/Forms: In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.
Isolation: Isolation date: 2000
Applications: 127TAg細胞DNA repair studies
Age: 14.5 days gestation embryo
Comments: 127TAg (Pms2 null [-/-]) is a mouse embryonic fibroblast (MEF) cell line derived from Pms-2 null embryos at day 14.5 of gestation. The Pms-2 deficient mice were constructed using ES-D3 [D3] embryonic stem cells [PubMed: 9096356]. 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).
Propagation: ATCC 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%.
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0°C
Subculturing: Protocol:
Remove and discard culture medium.
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.
127TAg細胞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.
Add appropriate aliquots of the cell suspension to new culture vessels.
An inoculum of 3 X 10(3) to 1 X 10(4) viable cells/cm2 is recommended.
Incubate cultures at 37?C.
Interval: Maintain cultures at a cell concentration between 3 X 10(3) and 1 X 10(4) cells/cm2.
Subcultivation Ratio: A subcultivation ratio of 1:6 to 1:8 is recommended
Medium Renewal: Every two to three days
Preservation: Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Doubling Time: 17 hours
Related Products: Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2002
recommended serum:ATCC 30-2020
References: 38299: 127TAg細胞Sobol RW, et al. Requirement of mammalian DNA polymerase-beta in base-excision repair. Nature 379: 183-186, 1996. PubMed: 8538772
88892: Sobol RW, et al. Base excision repair intermediates induce p53-independent cytotoxic and genotoxic responses. J. Biol. Chem. 278: 39951-39959, 2003. PubMed: 12882965
89411: Narayanan L, et al. Elevated levels of mutation in multiple tissues of mice deficient in the DNA mismatch repair gene Pms2. Proc. Natl. Acad. Sci. USA 94: 122-127, 1997. PubMed: 9096356
89413: 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
89432: Sobol RW, et al. The lyase activity of the DNA repair protein beta-polymerase protects from DNA-damage-induced cytotoxicity. Nature 405: 807-810, 2000. PubMed: 10866204
