Ku80+/+ (ATCC® CRL-3262)

Organism: Mus musculus, mouse  /  Cell Type: fibroblast  /  Tissue: embryo  / 

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Organism Mus musculus, mouse
Tissue embryo
Cell Type fibroblast
Product Format frozen 1.0 mL
Culture Properties adherent
Biosafety Level 1

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 12.5 days
Storage Conditions liquid nitrogen vapor phase
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Comments

Ku80+/+ cell line is wild type for Ku80. When paired with the mutant Ku80-/-, it can be used in cancer drug discovery and to investigate the role of DNA double-strand break repair in chromosomal aberrations.

Together, Ku790 and Ku80 make up the Ku heterodimer, which binds DNA double-strand break ends and is required for the non-homologous end joining(NHEJ) pathway of DNA repair. It is also required for V(D)J recombination, which utilizes the NHEJ pathway to promote antigen diversity in the mammalian immune system.

These cells were immortalized by SV40

Complete Growth Medium

The base medium for this cell line is DMEM (ATCC 30-2002). To make the complete medium add the following components to 500 mL of the base medium:

  • 56 mL fetal bovine serum (FBS; ATCC 30-2020)
Note: Be sure to heat inactivate the serum at 56° for 30 minutes
Subculturing
Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. Corning® T-75 flasks (catalog #430641) are recommended for subculturing this product.
  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with DPBS (ATCC catalog # 30-2200) to remove all traces of serum that contains trypsin inhibitor.
  3. Add 2.0 to 3.0 mL 0.25% (w/v) Trypsin- 0.53 mM 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.
    Cultures can be established between 2.0 x 104 and 4.0 x 105 viable cells/cm2.
  6. Incubate cultures at 37°C.
Interval: Maintain cultures at a cell concentration between 2.0 X 104 and 7.0 X 105 cell/cm2.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:8 is recommended
Medium Renewal: 2 to 3 times per week
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
Cells per Vial Approximately 2.0 to 3.0 x 106 cells
Volume 1.0 mL
Sterility Tests Bacteria and yeast: No growth
Mycoplasma: No growth
Viability ≥ 50%
Name of Depositor A Nussenzweig, LGI/NCI, Bethesda, MD
Year of Deposit 2002
References

Difillippantonio MJ, et al. DNA repair protein Ku80 suppresses chromosomal aberrations and malignant transformation. Nature 404(6777):510-514, 2000. PubMed: 10761921

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

Difillippantonio MJ, et al. DNA repair protein Ku80 suppresses chromosomal aberrations and malignant transformation. Nature 404(6777):510-514, 2000. PubMed: 10761921