Description: Cell line derived from an adenocarcinoma of the transverse colon

Description Key Words: Human adenocarcinoma transverse colon

Also Known As: No other names

Organism: Human (Homo sapiens)

Strain: N/A

Tissue: Colon

Growth Properties: Grows as loose aggregates of small rounded cells, some adherent, others in suspension.

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:1:8-1:16 Seeding density 1 x10⁴ cells/cm². Split sub confluent flasks (70-80%). Harvest the cells using 0.05% Trypsin/EDTA at 37°C for 5 minutes.

Culture Conditions: Cells maintained at 37°C and 5% CO₂.

Cryoprotectant Medium: 10% DMSO + 90% FCS

Additional Information: HNPCC patient

Mutations in:

–  p53 (wt/stop at aa 49)]

– beta-catenin (delta aa A5 to A76)

– B-Raf (aa V600E)

MSI

A33 negative

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_4443