Description: LIM 1899 is a columnar cell adenocarcinoma of the colon. Description Key Words: Columnar cell adenocarcinoma colon Also Known As: No other names Organism: Human (Homo sapiens) Strain: N/A Tissue: Colon Growth Properties: Adherent Morphology: Epithelial. Cells are small rounded and grow in clusters. Growth Medium: RPMI 1640 (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. Subcultivation Ratio: 1:8, Seeding density 1x104 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 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: Sporadic CRC. Mutated beta-catenin and K-Ras. A33 +ve.Medium:Growth Condition:BioVector NTCC Inc.www.biovector.net