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

Medium:
Growth Condition:

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