WM-266-4 (ATCC® CRL-1676)

Organism: Homo sapiens, human  /  Cell Type: Melanoma  /  Tissue: derived from metastatic site: skin  /  Disease: melanoma

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Organism Homo sapiens, human
Tissue derived from metastatic site: skin
Cell Type Melanoma
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 1
Disease melanoma
Age 58 years
Gender female
Storage Conditions liquid nitrogen vapor phase
Karyotype The cell line shares many properties with the parent tumor line including same karyology. This is a hypertriploid human cell line. The modal chromosome number is 82, occurring at 32%, with polyploidy at 8.6%. (Cells with 80 chromosome counts also occurred at a high frequency.) There were two paired [der(1)t(1;?) (p32;?), and der(11)t(2;11) (q22.1;p13)], and three to four single markers consistently found in each cell. Several other markers were also found in some cells only, including a small metacentric chromosome of unknown origin. Neither DMs nor HSRs were detected. Among the normal chromosomes, N11 had only single copy, and N1, N6, N8, N9, and N16 had two copies each. As expected, other normal chromosomes had three to four copies per cell. Two X chromosomes were generally found in each cell.
Derivation
Another cell line derived from the primary tumor of the same patient also is available (ATCC CRL-1675).
The WM-266-4 cell line was derived from a metastatic site of a malignant melanoma.
Genes Expressed
proteoglycan antigen characteristic of melanoma
Tumorigenic Yes
Effects
Yes, tumors developed within 21 days at 100% frequency (5/5) in nude mice inoculated subcutaneously with 107 cells.
Complete Growth Medium The base medium for this cell line is ATCC-formulated Eagle's Minimum Essential Medium, Catalog No. 30-2003. 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.
  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 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 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 1:3 to 1:6 is recommended
Medium Renewal: 2 to 3 times per week

Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.

Cryopreservation Complete growth medium, 95%; DMSO, 5%. Cell culture tested DMSO is available as ATCC Catalog No. 4-X.
Culture Conditions
Temperature: 37°C
STR Profile
Amelogenin: X
CSF1PO: 12
D13S317: 12,13
D16S539: 11,12
D5S818: 13
D7S820: 8
THO1: 7,9
TPOX: 8,11
vWA: 15,17
Name of Depositor M Herlyn
References

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.

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

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.