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: 1:4 – 1:8. Seeding density 0.8 x 10⁴ cells/cm². Spilt sub-confluent flasks (70-80%). 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:

• Homozygous deletion p14ARF
• Homozygous deletion p16INK4A
• V600E BRAF

Depositor: Peter Parsons – Queensland Institute of Medical Research Australia

References:

Parsons P.G, Morrison L. Melphalan-induced chromosome damage in sensitive and resistant human melanoma cell lines. International Journal of Cancer 21:(4) 407 – 522 , 1978. PubMed: 669848.

Castellano M et al.CDKN2A/p16 Is Inactivated in Most Melanoma Cell Lines. Cancer Research 57: 4868-4875. November 1. 1997. PubMed: 9354451.

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

Stark M. and Hayward N.Genome-Wide Loss of Heterozygosity and Copy Number Analysis in Melanoma Using High-Density Single-Nucleotide Polymorphism Arrays. Cancer Research 67: (6).2632-2642 2007. PubMed: 17363583.

Cellosaurus: CVCL_2604