Clone M-3 [Cloudman S91 melanoma] (ATCC® CCL-53.1)

Organism: Mus musculus, mouse  /  Cell Type: melanocyte  /  Tissue: skin  /  Disease: melanoma

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Organism Mus musculus, mouse
Tissue skin
Cell Type melanocyte
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 1
Disease melanoma
Gender male
Strain DBA
Storage Conditions liquid nitrogen vapor phase
Karyotype Stemline number is hypertetraploid. Karyotype is stable within stemline number. Marker chromosomes: A medium size chromosome with a submedian centromere and a smaller chromosome with a median centromere., The remaining 81 chromosomes have terminal centromeres, the first one is larger than normal. A minute chromosome was noted in 20% of the cells.
Derivation
Clone M-3, a melanin-producing cell line was adapted to cell culture by Y. Yasumura, A.H. Tashjian and G. Sato from a Cloudman S91 melanoma in a (C X DBA) F1 male mouse obtained from the Jackson Memorial Laboratory, Bar Harbor, Maine.
Clinical Data
male
Genes Expressed
melanin
Cellular Products
melanin
Tumorigenic Yes
Effects
Yes, in C57BL mice
Virus Susceptibility Herpes simplex virus
Vaccinia virus
Pseudorabies virus
Vesicular stomatitis, Glasgow (Indiana)
Vesicular stomatitis, Orsay (Indiana)
Virus Resistance
poliovirus 1
Comments

Tested and found negative for ectromelia virus (mousepox).

Complete Growth Medium The base medium for this cell line is ATCC-formulated F-12K Medium, Catalog No. 30-2004. To make the complete growth medium, add the following components to the base medium:
  • fetal bovine serum to a final concentration of 2.5%
  • horse serum to a final concentration of 15%
  • 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.

    1.  Remove and discard culture medium.
    2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin 0.03% (w/v) EDTA solution to remove all traces of serum which 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 37oC 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 into new culture vessels.
    6. Incubate cultures at 37°C.

    Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:4 is recommended

    Medium Renewal: Every other day

    Cryopreservation
    culture medium 95%; DMSO, 5%
    Culture Conditions
    Temperature: 37°C
    Atmosphere: air, 95%; carbon dioxide (CO2), 5%
    Name of Depositor G Sato
    References

    Yasamura Y, et al. Establishment of four functional, clonal strains of animal cells in culture. Science 154: 1186-1189, 1966. PubMed: 4288399

    Schmidt W, et al. Cell-free tumor antigen peptide-based cancer vaccines. Proc. Natl. Acad. Sci. USA 94: 3262-3267, 1997. PubMed: 9096381

    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
    References

    Yasamura Y, et al. Establishment of four functional, clonal strains of animal cells in culture. Science 154: 1186-1189, 1966. PubMed: 4288399

    Schmidt W, et al. Cell-free tumor antigen peptide-based cancer vaccines. Proc. Natl. Acad. Sci. USA 94: 3262-3267, 1997. PubMed: 9096381