Description: NZM5 is a semi-adherent cell line derived from a 79-year-old male patient with metastatic melanoma (axillary lymph node).

Description Key Words: Human metastatic melanoma 

Also Known As: No other names

Organism: Human (Homo sapiens) 

Tissue: Melanoma metastatic to axillary lymph node

Growth Properties: Semi-Adherent the cells grow as a loosely adherent monolayer and also in suspension

Doubling Time: 48 hours

Morphology: Epithelial and rounded in suspension

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: 2-3 times per week.

Subcultivation Ratio: 1:5, Seeding density 4.0 x10⁴cells/cm². Split subconfluent cultures (70-80%). Harvest the cells using 0.05% Trypsin/EDTA at 37°C for 2 minutes. Cells may take 2-3 days to seed after thaw. They are 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: 

• Wild-type for TP53, PIK3CA, CDKN2A/B, PTEN, NRAS, BRAF
• Close to diploid: DNA content 40-43 chromosomes 
• Resistant to temozolomide 
• MGMT expressed.

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

Reference:

Original Reference:

Marshall ES, Matthews JHL, Shaw JHF, Nixon J, Tumewu P, Finlay GJ, Holdaway KM, Baguley BC.

Radiosensitivity testing of human melanoma cell lines: comparison of [3H]-thymidine incorporation and soft agar clonogenic assays.

Eur J Cancer 1994; 30A: 1370-1376.

PubMed 7999427

Other Reference:

Parmar J, Marshall ES, Charters GA, Holdaway KM, Shelling AN, Baguley BC.

Radiation-induced cell cycle delays and p53 status of early passage melanoma lines. Oncol Res 2000; 12: 149-155. PubMed 11216673

Cellosaurus: CVCL_D837