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Datasheet Protocol

WM266-4 Viable Cells

5 References
WM266-4-01-0001 WM266-4-01-0005 WM266-4-01-0010
1 million cells 5 x 1 million cells 10 x 1 million cells
Frozen Cell Suspension Frozen Cell Suspension Frozen Cell Suspension
ELISA, IHC, Cellular Assay, Other
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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Background

WM266-4 is a metastatic human melanoma cell line with small flat mesenchymal morphology. This cell line was derived from the same patient as the cell lines WM115, WM239A, and WM165-1. WM115 cell line originated from the primary tumor, and WM266-4, WM165-1 and WM239A were from individual lymph-node metastases. The subject (55-year-old female) displayed VGP with a Clark level III tumor with thickness of 2.24mm.This cell line features the specific V600D (Val600Asp) mutation at codon 600 in the BRAF gene. This cell line also expresses PTEN loss of function including hemizygous deletion. WM266-4 cell line is wild type for N-RAS, c-KIT, and CDK4 genes. WM266-4 cells produce xenograft tumors when injected into immunocompromised mice.

Product Details

WM266-4 Viable Cells - WM266-4-01-0001
Melanoma cells, patient derived tumor cells, tumor models, skin cancer testing, xenograft production, Human melanoma tumor cell
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

Cellular Assay, ELISA, IHC, Other  - View References
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
small flat mesenchymal
V600D
WT
WT
Hemizygous Deletion
Tumor Specialized Media with 2% HI-FBS
Cells should be maintained between 30 – 95% confluence in tumor specialized medium with 2% FBS; split cultures 1:3 every 7 days 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.
(). Cytokine and Chemokine Receptor Patterns of Human Malignant Melanoma Cell Lines. Int J Mol Sci.
Applications
RNA seq, qPCR, Reverse Transcription, RT-PCR Analysis; Statistical Analysis; In Vitro Assay
(). ---Inhibiting FAK-Paxillin Interaction Reduces Migration and Invadopodia-Mediated Matrix Degradation in Metastatic Melanoma Cells. Cancers (Basel).
Applications
IF, Confocal Microscopy; WB, IB, PCA
(). Epigenetic silencing of CDR1as drives IGF2BP3-mediated melanoma invasion and metastasis. Cancer Cell.
Applications
Other
(). Melanoma Progression Inhibits Pluripotency and Differentiation of Melanoma-Derived iPSCs Produces Cells with Neural-like Mixed Dysplastic Phenotype. Stem Cell Reports.
Applications
Cell Proliferation Assay
(). Targeting the Proteasome-Associated Deubiquitinating Enzyme USP14 Impairs Melanoma Cell Survival and Overcomes Resistance to MAPK-Targeting Therapies. Mol Cancer Ther.
Applications
EIA, ELISA

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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