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 0.6 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:

• Homozygous deletion CDNK2A
• V600E BRAF

Depositor: Peter Parsons – Queensland Institute of Medical Research Australia

References:

Maynard, K. and Parsons P.G. Cross-sensitivity to methylating agents, hydroxyurea and methotrexate in human tumour cells of the Mer- phenotype. Cancer Research: 46:5009-5013, 1986. PubMed: 3756862.

Cellosaurus: CVCL_2605