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).

Medium:
Growth Condition:

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