ATCC® Number: CRL-1676™ Price: $318.00
Designations: WM-266-4
Depositors: M Herlyn
Biosafety Level: 1
Shipped: frozen
Medium & Serum: See Propagation
Growth Properties: adherent
Organism: Homo sapiens （human）
Morphology: epithelial
Source: Organ: skin
Disease: melanoma
Cellular Products: proteoglycan antigen characteristic of melanoma
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Tumorigenic: Yes
DNA Profile （STR）: Amelogenin: X
CSF1PO: 12
D13S317: 12,13
D16S539: 11,12
D5S818: 13
D7S820: 8
THO1: 7,9
TPOX: 8,11
vWA: 15,17
Cytogenetic Analysis: 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.
Age: 58 years
Gender: female
Comments: The WM-266-4 cell line was derived from a metastatic site of a malignant melanoma.
Another cell line derived from the primary tumor of the same patient also is available （ATCC CRL-1675）.
Propagation: ATCC 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%.
Temperature: 37.0°C
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 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.
Subc*tion Ratio: A subc*tion ratio of 1:3 to 1:6 is recommended
Medium Renewal: 2 to 3 times per week
Preservation: Freeze medium: Complete growth medium, 95%; DMSO, 5%
Storage temperature: liquid nitrogen vapor phase
Related Products: Recommended medium （without the additional supplements or serum described under ATCC Medium）:ATCC 30-2003
recommended serum:ATCC 30-2020
derived from same individual:ATCC CRL-1675
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