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: MM537 requires growth medium to be changed 3 times each week.

Subcultivation Ratio: 1:4. When seeded at 2 x 10⁶ cells/T75 flask the cells will need to be passaged every 5 days. Split sub-confluent flasks (70-80% confluent). Flasks do not reach confluence. Harvest the cells using 0.05% Trypsin/EDTA at 37°C for 5 minutes. The cells are sensitive to trypsin so exposure time to trypsin should be kept to a minimum. The cell line takes 2-3 days to recover from trypsinisation.

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 CDKN2A
• 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.

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.

Celloasurus: CVCL_4451