Description: NZM12 is an adherent cell line derived from a male 19yr old patient with melanoma metastatic to small intestine.

Description Key Words: Human metastatic melanoma to small intestine 

Also Known As: No other names

Organism: Human (Homo sapiens) 

Tissue: Melanoma metastatic to small intestine

Growth Properties: Adherent

Doubling Time: 40-42 hours

Morphology: small adherent and star shaped

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. 

Note: the NZM12 cells can take 48 hours to seed after thaw or passaging.

Subculturing Procedure:

Medium Renewal: 2-3 times per week.

Subcultivation Ratio: 1:4 – 1:8 Seeding density 0.9 – 1.3 x10⁴cells/cm². Split subconfluent cultures(70-80%). Harvest the cells using 0.05% Trypsin/EDTA at 37°C for 5 minutes. Cells take 48 hours to seed.

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 PIK3CA, PTEN, NRAS
• Partial deletion CDKN2A/B
• BRAF mutation V600E
• Resistant to temozolomide
• MGMT expressed 

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

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