Description: MDCK cell line (ECACC catalogue no. 84121903) adapted to grow in the protein-free medium 'MDCK Protein-free Medium' (Sigma 14581C). Stirred culture (or Ultra low attachment flasks Corning 3814) is recommended, cells grow in suspended clumps.

Also Known As:

Species: Dog

Tissue: Kidney

Growth Properties: Aggregates in suspension

Morphology:

Growth Medium: MDCK Protein-free medium (Sigma 14581C) + 3.3mM Glutamine. When freezing cells down use Cell Freezing Medium (Sigma C2639) + 10% DMSO or use 50:50 fresh culture medium:conditioned medium plus 10% DMSO.

Subculturing Procedure: Stirred culture (or Ultra low attachment flasks Corning 3814) is recommended, cells grow in suspended clumps.Viability may be poor on resuscitation and may initially decrease further. Full recovery may take up to 2 weeks. A centrifugation step to remove the cryoprotectant is essential. Rapidly thaw the frozen ampoule in a water bath at 37°C for 1-2 minutes. Transfer the contents to a centrifuge tube and slowly add 5-10ml of pre-warmed growth media. Remove a sample for counting. Centrifuge at 100g for 2-3 minutes to pellet cells and seed at a relatively high density of 5-7 x 100,000 cells/ml. Leave culture flask upright and observe regularly until viable proliferating cells are seen. Once established use a split ratio of 1:2 approximately every 4 to 5 days; 5% CO2; 37°C.

Release Conditions: No

Depositor: European Collection of Cell Cultures, Health Protection Agency, Porton Down, Salisbury, SP4 0JG, UK

Originator: Yes

References: None Specified By Depositor

Hyperlink to ECACC Cell Line Data Sheet: 02050101