Description: Colon carcinoma

Description Key Words: Human colon carcinoma

Also Known As: No other names

Organism: Human (Homo sapiens)

Strain: N/A

Tissue: Colon

Growth Properties: Cell line grows as floating and loose aggregates with some adherent cells.

Morphology: Epithelial

Growth Medium: RPMI1640 (with 2mM L-Glutamine + 25mM HEPES) + 10% Foetal Calf Serum (FCS) + Insulin 0.6μg/ml, Hydrocortisone 1μg/ml, 1-Thioglycerol 10μM.

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 each week. Feed and passage as for semi-adherent culture (collect medium with floating cells and then trypsinise the adherent population pool both together before reseeding).

Subcultivation Ratio: 1:4 – 1:8 Seeding density 1 – 2 x10⁴ cells/cm². Split sub-confluent cultures (70-80%). Harvest the cells using 0.05% Trypsin/EDTA at 37°C for 5 minutes. Cells may take 2-3 days to seed after thawing or trypsinisation. 

Culture Conditions: Cells maintained at 37°C and 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: Derived from colon carcinoma, adherent and loose aggregates

MSI, A33 negative, truncated APC (stop at aa 1544)

Mutated p53 (aa G245D)

Heterozygous mutated K-Ras (aa Q61H)

Mutated PI3K (aa H1047R p110alpha).

Depositor: Professor Tony Burgess – Ludwig Institute for Cancer Research Ltd, Melbourne, Australia

Reference:

Zhang H. et al.

Selective inhibition of proliferation in colorectal carcinoma cell lines expressing mutant APC or activated B-Raf

Int.J.Cancer 2009 July15; 125(2):297-307

PubMed 19378335

Cellosaurus: CVCL_4442