MB355 (ATCC® CRL-2818)

Organism: Mus musculus, mouse  /  Cell Type: fibroblast, spontaneously immortalized  / 

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Organism Mus musculus, mouse
Cell Type fibroblast, spontaneously immortalized
Product Format frozen
Morphology fibroblast
Culture Properties adherent
Biosafety Level 1
Age 14.5 days gestation embryo
Applications
DNA repair studies
Storage Conditions liquid nitrogen vapor phase
Images
Derivation
MB355 (Polb/p53 double null) is a mouse embryonic fibroblast (MEF) cell line derived from polymerase (DNA directed), beta (Polb)/p53 double null embryos at day 14.5 of gestation. The cells were spontaneously immortalized at passage 5 due to the loss of the p53 gene. The cells are transgenic for lambda LIZ (Lac I/cII).
Comments
MB355 (Polb/p53 double null) is a mouse embryonic fibroblast (MEF) cell line derived from polymerase (DNA directed), beta (Polb)/p53 double null embryos at day 14.5 of gestation. The cells were spontaneously immortalized at passage 5 due to the loss of the p53 gene. 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%.
Subculturing
Protocol:
  1. Remove and discard culture medium.
  2. 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.
  3. 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.
  4. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
  5. Add appropriate aliquots of the cell suspension to new culture vessels.
    An inoculum of 2 to 6 X 10(3) viable cells/cm2 is recommended.
  6. 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: Every two to three days
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 20 hours
Name of Depositor RW Sobol
Passage History
MB355 (Polb/p53 double null) is a mouse embryonic fibroblast (MEF) cell line derived from polymerase (DNA directed), beta (Polb)/p53 double null embryos at day 14.5 of gestation. The cells were spontaneously immortalized at passage 5 due to the loss of the p53 gene. The cells are transgenic for lambda LIZ (Lac I/cII).
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

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

Jacks T, et al. Tumor spectrum analysis in p53-mutant mice. Curr. Biol. 4: 1-7, 1994. PubMed: 7922305

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

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
Other Documentation
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

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

Jacks T, et al. Tumor spectrum analysis in p53-mutant mice. Curr. Biol. 4: 1-7, 1994. PubMed: 7922305

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