CRL-1676 WM-266-4 人黑素瘤細(xì)胞

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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CRL-1676 WM-266-4 人黑素瘤細(xì)胞

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