ATCC® Number: CCL-34™
Designations: MDCK (NBL-2)
Depositors: S Madin, NB Darby
Biosafety Level: 1
Shipped: frozen
Medium & Serum: See Propagation
Growth Properties: adherent
Organism: Canis familiaris
Morphology: epithelial
Source: Organ: kidney
Disease: normal
Cellular Products: keratin
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Isolation: Isolation date: September, 1958
Applications: transfection host (Nucleofection technology from Lonza
Roche FuGENE® Transfection Reagents)
Virus Susceptibility: Human Coxsackievirus B 5
Reovirus type 2
Adeno-associated virus 4
Vaccinia virus
Vesicular stomatitis virus
Adeno-associated virus 5
Human Coxsackievirus B 3
Human Coxsackievirus B 4
Human poliovirus 2
Reverse Transcript: negative
Cytogenetic Analysis: Polyploidy 0.2%. Two large submetacentric chromosomes noted, presumably X chromosomes, and one or two additional chromosomes with median or submedian centromeres.
Age: adult
Gender: female
Comments: The MDCK cell line was derived from a kidney of an apparently normal adult female cocker spaniel, September, 1958, by S.H. Madin and N.B. Darby.The cells are positive for keratin by immunoperoxidase staining.MDCK cells have been used to study processing of beta amyloid precursor protein and sorting of its proteolytic products.
Propagation: ATCC 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%.
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0°C
Subculturing: Protocol:
Remove and discard culture medium.
Rinse the cell layer twice with 0.25% (w/v) Trypsin - 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
Add 2.0 to 3.0 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.
Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:6 is recommended
Medium Renewal: Every 2 to 3 days
Preservation: Freeze medium: Complete growth medium 95%; DMSO, 5%
Storage temperature: liquid nitrogen vapor phase
Related Products: Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2003
recommended serum:ATCC 30-2020
parental cell line:ATCC CCL-34.2
0.25% (w/v) Trypsin - 0.53 mM EDTA in Hank' BSS (w/o Ca++, Mg++):ATCC 30-2101
Cell culture tested DMSO:ATCC 4-X
References: 18385: Didier ES, et al. Characterization of Encephalitozoon (Septata) intestinailis isolates cultured from nasal mucosa and bronchoalveolar lavage fluids of two AIDS patients. J. Eukaryot. Microbiol. 43: 34-43, 1996. PubMed: 8563708
22808: Haass C, et al. Polarized sorting of beta-amyloid precursor protein and its proteolytic products in MDCK cells is regulated by two independent signals. J. Cell Biol. 128: 537-547, 1995. PubMed: 7860629
25972: Gaush CR, et al. Characterization of an established line of canine kidney cells (MDCK). Proc. Soc. Exp. Biol. Med. 122: 931-935, 1966. PubMed: 5918973
28301: Loffler S, et al. CD9, a tetraspan transmembrane protein, renders cells susceptible to canine distemper virus. J. Virol. 71: 42-49, 1997. PubMed: 8985321
32843: Mead JR, et al. In vitro expression of mRNA coding for a Cryptosporidium parvum oocyst wall protein. J. Eukaryot. Microbiol. 43: 84-85, 1996. PubMed: 8822876
32899: von Dippe P, et al. The functional expression of sodium-dependent bile acid transport in Madin-Darby canine kidney cells transfected with the cDNA for microsomal epoxide hydrolase. J. Biol. Chem. 271: 18176-18180, 1996. PubMed: 8663355
33046: Panneerselvam K, Freeze HH. Mannose enters mammalian cells using a specific transporter that is insensitive to glucose. J. Biol. Chem. 271: 9417-9421, 1996. PubMed: 8621609
33080: Stuart RO, et al. Dependence of epithelial intercellular junction biogenesis on thapsigargin-sensitive intracellular calcium stores. J. Biol. Chem. 271: 13636-13641, 1996. PubMed: 8662885
33127: Grindstaff KK, et al. Translational regulation of Na,K-ATPase alpha1 and beta1 polypeptide expression in epithelial cells. J. Biol. Chem. 271: 23211-23221, 1996. PubMed: 8798517
