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WM989 Viable Cells

WM989-01-0001 WM989-01-0005 WM989-01-0010
1 million cells 5 x 1 million cells 10 x 1 million cells
Frozen Cell Suspension Frozen Cell Suspension Frozen Cell Suspension
Human
$1,089.00 /Per Item
$3,629.00 /Per Item
$5,445.00 /Per Item
1 million cells $1,089.00
5 x 1 million cells $3,629.00
10 x 1 million cells $5,445.00
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Product Details

WM989 Viable Cells - WM989-01-0001
Melanoma, patient derived tumor, tumor models, skin cancer, xenograft
Human

Target Details

Cells are sterile, validated by short tandem repeat profiling, and are tested as negative for mycoplasma. It is recommended that cell lines are tested for mycoplasma contamination and short tandem repeat (STR) profiling every 10 passages or each time a frozen seed stock is made. See cell culture protocol for additional details.

Application Details

The key applications of these cell lines include genetic studies, xenograft production, drug testing, and drug target discovery. These cell line models can be used in various biological assays, and for identifying critical target genes, and cell signaling pathways.

Cell Line Data

Human Melanoma
Viable Cells
V600E
WT
WT
ND
Tumor Specialized Media with 2% HI-FBS
Cells should be maintained between 30 – 95% confluence in tumor specialized medium with 2% FBS; split cultures 1:10 every 1 week using 0.25% trypsin/EDTA.
36°C with 5% CO2

Formulation

1.0 million cells/mL Count By Hemocytometer
None
None
None

Shipping & Handling

Dry Ice
Cells are frozen with 90% FBS/10% DMSO solution at about 1x10^6 cells/ml. Store vial in liquid nitrogen upon arrival.
Expiration date is two (2) years from date of receipt.
WM989 is a melanoma cell line. This cell line features the specific V600E (Val600Glu) mutation at codon 600 in the BRAF gene. This mutation causes constitutively active kinase activity and activation of MEK and ERK signaling pathway. This cell line expresses wild type N-RAS, c-KIT, and CDK4 genes. WM989 cells produce xenograft tumors when injected into immunocompromised mice.

How can I ensure that the cells will be viable after resuspension?

Complete adherence to the protocol is necessary to ensure the viability of the cells. Cells should be maintained between 30-95% confluence in Tumor Specialized media with 2% FBS (heat inactivated) unless otherwise noted in cell line specific instructions.

How should I handle the cells before culturing?

Before thawing, frozen cells can be stored in liquid nitrogen or at -80°C only. To thaw the melanoma cells, move the vial from liquid nitrogen to dry ice immediately. Melanoma cells should be thawed quickly. Freeze thaw cycles can damage / kill the cells and should be avoided. Centrifuge the cells at 1500 rpm (500 x g) for 5 minutes at room temperature. Discard the supernatant. Resuspend cell pellet in 5 mL of Tumor Specialized Media with FBS (heat inactivated) and transfer to a T-25 flask. Place the flask in a humidified incubator (5% CO2) at 36°C overnight. Check the flask after 24 hours for attachment of cells to the flask. If cells are attached, remove the media and add fresh media to the flask.

Which media should I use to grow melanoma cells?

Tumor specialized Media with 2% FBS is preferred media for melanoma cell lines. However, alternative media (not preferred) recommendations include: 1. DMEM with 5% FBS; 2. RPMI with 5% FBS.

Do you have recommended sources for Tumor Specialized Media components?

The recommended manufacturer for each component of the Tumor Specialized Media are: MCDB-153 (Sigma-Aldrich, item# M-7403), Leibovitz’s L-15 (Sigma-Aldrich, item# L1518), FBS (heat inactivated) (Rockland, item# FBS-010-0100), Calcium Chloride (Sigma-Aldrich, item# C5670), Sodium Bicarbonate [Added to MCDB-153 preparation] (Sigma-Aldrich, item# S5761). Please consult the protocol for detailed instructions on the preparation of the media.

What is the optimal growth temperature for melanoma cells?

Melanoma cells grow best at 36°C (not 37°C) with 5% CO2. These incubation conditions must be followed exactly to ensure the viability and growth of the cells.

What is the recommended media for freezing viable melanoma cells?

Freezing media containing 90% FBS and 10% DMSO. Thawed cells should not be frozen again. Melanoma cells should be frozen slowly but thawed quickly.

What is the source of the melanoma cell lines?

The melanoma cell lines were established from patient-derived tumors. Information about the patients or treatments received is not available.

Are melanoma cell lines immortalized?

Melanoma cell lines are considered immortalized because they can replicate indefinitely.

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