127TAg細(xì)胞

生長(zhǎng)狀態(tài)： 貼壁生長(zhǎng)

運(yùn)輸方式： 凍存運(yùn)輸

細(xì)胞形態(tài)： 成纖維樣

細(xì)胞類型： 其他細(xì)胞類型

是否是腫瘤細(xì)胞： 0

物種來源： 小鼠

ATCC Number： CRL-2817?

數(shù)量： 大量

年限： 14.5 days gestation embryo

器官來源： 胚胎

Designations: 127TAg

127TAg細(xì)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(xì)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(xì)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(xì)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