Cell Line Description: Melanoma cell line. From metastatic site: Lymph node.

Also Known As:

Organism: Human (Homo sapiens)

Tissue: Skin

Growth Properties: Adherent

Morphology: Epithelial

Growth Medium: RPMI 1640 (with 2mM L-Glutamine + 25mM HEPES) + 10% Foetal Calf Serum (FCS)

Resuscitation: Remove protective cryoflex layer around the ampoule prior to thawing. Thaw the ampoule by gently agitating in a 37°C waterbath; thawing should be rapid (around 2 minutes). A centrifugation step to remove the cryoprotectant after thawing is necessary for this cell line.

Subculturing Procedure:

Medium Renewal:

Subcultivation Ratio: 1:8. Seeding density 1.2 x 10⁴ cells/cm². Harvest the cells using 0.05% Trypsin/EDTA at 37°C for 5 minutes.

Culture Conditions: Cells incubated at 37°C with 5% CO2.

Cryoprotectant Medium: 10% DMSO + 90% FCS

Handling Procedure for Frozen Cells: Upon receipt, frozen ampoules should be transferred directly to liquid nitrogen storage without delay, if not to be used immediately. Storage at -80°C may result in loss of viability.

Additional Information:

Mutations:

• V600E BRAF

Depositor: Peter Parsons – Queensland Institute of Medical Research Australia

References:

Castellano M et al. CDKN2A/p16 Is Inactivated in Most Melanoma Cell Lines. Cancer Research: 57: 4868-4875 1997. PubMed 9354451.

Pavey S et al. Microarray expression profiling in melanoma reveals a BRAF mutation signature, Oncogene 23: 4060–4067, 2004. PubMed: 15048078.

Packer L. et al. Osteopontin is a downstream effector of the PI3-kinase pathway in melanomas that is inversely correlated with functional PTEN, Carcinogenesis 27: (9) 1778-1786, 2006. PubMed: 16571650.

Cellosaurus: CVCL_2606