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Permits and Restrictions	View Permits
Organism	Canis familiaris, dog
Tissue	kidney; distal tubule
Product Format	frozen
Morphology	epithelial-like
Culture Properties	adherent
Biosafety Level	1 Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
Disease	normal
Age	adult
Storage Conditions	liquid nitrogen vapor phase

Karyotype	hyperdiploid canine cell line with a modal chromosome number of 91 with low polyploidy rate. Several unidentifiable marker chromosomes were present in most of the cells examined.
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Derivation	Cell line was derived by cloning (limited dilution) the parental cell line MDCK (ATCC CCL-34).
Antigen Expression	E-cadherin (epithelial cell adhesion molecule), expressed Zona Occludens (ZO-1) (tight junction protein), expressed fibroblast-specific protein (FSP), not expressed cytokeratin (CK1, 4, 5, 6, 8, 10, 13, 18, 19),expressed sialic receptors: alpha 2,3-galactose (avian)and alpha 2,6-galactose (human); expressed
Genes Expressed	Zona Occludens (ZO-1) (tight junction protein), expressed,E-cadherin (epithelial cell adhesion molecule), expressed,fibroblast-specific protein (FSP), not expressed ,cytokeratin (CK1, 4, 5, 6, 8, 10, 13, 18, 19),expressed,sialic receptors: alpha 2,3-galactose (avian)and alpha 2,6-galactose (human); expressed
Virus Susceptibility	Influenza A virus Influenza B virus
Comments	This cell line is susceptible to a wide range of influenza virus and is sensitive to epsilon toxin of C. perfringens.

Complete Growth Medium	The base medium for this cell line is ATCC-formulated Eagle's Minimum Essential Medium, Catalog No. 30-2003. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
Subculturing	Volumes used in this protocol are for 75 cm²flasks; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. Remove and discard culture medium. Briefly rinse the cell layer with Ca++/Mg++ free Dulbecco's phosphate-buffered saline (D-PBS) or 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum which contains trypsin inhibitor. Add 1.0 to 2.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting. Add appropriate aliquots of the cell suspension to new culture vessels. An inoculum of 1 X 10⁴ to 2 X 10⁴ viable cells/cm² is recommended. Incubate cultures at 37°C. Subculture when the cell concentration is between 7 X 10⁴ and 1 X 10⁵ cells/cm². Subcultivation ratio: A subcultivation ratio of 1: 2 to 1:6 is recommended. Medium renewal: Every 2 to 3 days.
Cryopreservation	Freeze medium: Complete growth medium, 95%; DMSO, 5% liquid nitrogen vapor phase
Culture Conditions	Temperature: 37°C Atmosphere: air, 95%; carbon dioxide (CO₂), 5%

Population Doubling Time	approximately 22 hours

Name of Depositor	Y. Reid, E. Cedrone and E-Eckard-Amar, ATCC
Year of Origin	Jan 2007
References	Cedrone E, et al. Tissue-culture adapted Influenza virus strains. ATCC Connection volume 29 (2): 4-5 and 15, 2009.

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P: 1300 552 003

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