To ATCC Valued Customers,

ATCC stands ready to support our customers’ needs during the coronavirus pandemic. If you experience any issues with your products or services, please contact ATCC Customer Service at sales@atcc.org. For Technical questions please contact tech@atcc.org. Thank you.
X

Privacy Policy Update

We remain dedicated to protecting your data and experience throughout our platforms. We have updated our Privacy Policy and your continued use of the Site means you have accepted the revised Privacy Policy. View now >
X

B16-F10 (ATCC® CRL-6475)

Organism: Mus musculus, mouse  /  Tissue: skin  /  Disease: melanoma

Permits and Restrictions

View Permits

Organism Mus musculus, mouse
Tissue
skin
Product Format frozen
Morphology mixture of spindle-shaped and epithelial-like cells
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.

Disease melanoma
Strain C57BL/6J
Applications
This line is a suitable transfection host.
Storage Conditions liquid nitrogen vapor phase
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

Volumes are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally 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 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.
  6. Incubate cultures at 37°C.

Subcultivation Ratio: A subcultivation ratio of 1:10 is recommended

Medium Renewal: Every 2 to 3 days

Cryopreservation
Freeze medium: culture medium, 95%; DMSO, 5%
Storage temperature: liquid nitrogen vapor phase
Culture Conditions

Temperature: 37°C

Atmosphere: air, 95%; carbon dioxide (CO2), 5%

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
FAQ's
  1. Color of growth media for CRL-6475
    The cells produce melanin causing the culture medium to appear dark brown or black, especially when approaching a high level of confluence.
    Date Updated: 3/27/2014
  2. Gender of the CRL-6475, B16f-10 cells


    Date Updated: 8/2/2019

References

Fidler IJ. Biological behavior of malignant melanoma cells correlated to their survival in vivo. Cancer Res. 35: 218-224, 1975. PubMed: 1109790

Fidler IJ, et al. Tumoricidal properties of mouse macrophages activated with mediators from rat lymphocytes stimulated with concanavalin A. Cancer Res. 36: 3608-3615, 1976. PubMed: 953987

Fidler IJ, Bucana C. Mechanism of tumor cell resistance to lysis by syngeneic lymphocytes. Cancer Res. 37: 3945-3956, 1977. PubMed: 908034

Fidler IJ, Kripke ML. Metastasis results from preexisting variant cells within a malignant tumor. Science 197: 893-895, 1977. PubMed: 887927

Briles EB, Kornfeld S. Isolation and metastatic properties of detachment variants of B16 melanoma cells. J. Natl. Cancer Inst. 60: 1217-1222, 1978. PubMed: 418183

Fidler IJ. Selection of successive tumour lines for metastasis. Nat. New Biol. 242: 148-149, 1973. PubMed: 4512654

Li M, et al. Loss of intracisternal A-type retroviral particles in BL6 melanoma cells transfected with MHC class I genes. J.Gen. Virol. 77: 2757-2765, 1996. PubMed: 8922469