Description: NZM40 is an adherent cell line derived from a metastatic melanoma from a 47yr old female patient in 2004.

Description Key Words: Human metastatic melanoma 

Also Known As: No other names

Organism: Human (Homo sapiens) 

Growth Properties: Adherent. Cells are large and the yield per flask is low at around 2 million cells per T75 flask.

Doubling Time: 38 hours

Morphology: Epithelial

Growth Medium: This cell line was established by the depositor in: Alpha modified Minimum Essential Medium (MEM) supplemented with 5% Foetal Bovine Serum (FBS)”, 5 ug/ml Insulin, 5 ug/ml Transferrin, 5 ng/ml Sodium Selenite (ITS), in an atmosphere of 5%O₂, 5%CO₂ and 90%N₂.

At CellBank Australia it has been cultured in: Alpha modified Minimum Essential Medium (MEM) supplemented with 10% Foetal Bovine Serum (FBS)”, 5 ug/ml Insulin, 5 ug/ml Transferrin, 5 ng/ml Sodium Selenite (ITS), in an atmosphere of 5%CO₂.

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: 3 times per week.

Subcultivation Ratio: 1:4 – 1:8 Seeding density 0.7 x10⁴cells/cm². Split subconfluent cultures (70-80%). Harvest the cells using 0.05% Trypsin/EDTA at 37°C for 5 minutes. Note that this cell line is trypsin sensitive so care should be taken to minimise exposure time to trypsin.

Culture Conditions: Incubate the culture 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: 

• Aneuploid 1.82x and 3.06x diploid;
• Mutations: p53 mutant (del 249-253)”, NRAS mutant Q61H, PTEN, BRAF
• Wild-type for CDKN2A/B PIK3CA
• Temozolomide resistant; MGMT not expressed.

Depositor: Professor Bruce Baguley – The University of Auckland, New Zealand

Reference:

Original Reference:

Kim JE, Stones C, Joseph WR, Leung E, Finlay GJ, Shelling AN, Phillips WA, Shepherd PR, Baguley BC.

Comparison of growth factor signalling pathway utilisation in cultured normal melanocytes and melanoma cell lines.

BMC Cancer. 2012 Apr 4; 12:141. doi: 10.1186/1471-2407-12-141.

PubMed 22475322

Cellosaurus: CVCL_D832