MDCK (NBL-2) (ATCC® CCL-34)

Organism: Canis familiaris  /  Tissue: kidney  /  Disease: normal

Organism Canis familiaris
Tissue kidney
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 1
Disease normal
Age adult
Gender female
Applications
This cell line is a suitable transfection host and is useful for influenza research.
Storage Conditions liquid nitrogen vapor phase
Karyotype Modal chromosome number for species=78 Range 77-80; 87-90.
Images
Derivation
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.
Clinical Data
female

Genes Expressed
The cells are positive for keratin by immunoperoxidase staining.
Cellular Products
keratin
Virus Susceptibility Human Coxsackievirus B 5
Reovirus type 2
Adeno-associated virus 4
Vaccinia virus
Vesicular stomatitis virus
Adeno-associated virus 5
Human Coxsackievirus B3
Human Coxsackievirus B 4
Human poliovirus 2
Comments

This line is hyperdiploid and there is a bi-modal chromosome number distribution. There are no consistent identifiable marker chromosomes. One normal X chromosome is present in most spreads.

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.

 

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 are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes. Corning® T-75 flasks (catalog #430641) are recommended for subculturing this product.
  1. Remove and discard culture medium.
  2. 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.
  3. 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.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  5. Add appropriate aliquots of the cell suspension to new culture vessels.
  6. 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
Cryopreservation
Freeze medium: Complete growth medium 95%; DMSO, 5%
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
Name of Depositor S Madin, NB Darby
Year of Origin September, 1958
References

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

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

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

Loffler S, et al. CD9, a tetraspan transmembrane protein, renders cells susceptible to canine distemper virus. J. Virol. 71: 42-49, 1997. PubMed: 8985321

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

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

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

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

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

Cross References

Nucleotide (GenBank) : AB032025 Canis familiaris mRNA for ubiquitin, partial cds.

Nucleotide (GenBank) : D01020 Canis familiaris interleukin-8 gene, partial sequence.

Basic Documentation
Other Documentation
FAQ's
  1. Morphology of ATCC® CCL-34, MDCK


    Date Updated: 2/5/2013

  2. ATCC® CCL-34: Vacuoles in MDCK Cells

    Vacuoles in MDCK cells (ATCC® CCL-34 ™) are often a sign of stress. Some causes of stress include:


    Date Updated: 2/5/2013
References

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

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

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

Loffler S, et al. CD9, a tetraspan transmembrane protein, renders cells susceptible to canine distemper virus. J. Virol. 71: 42-49, 1997. PubMed: 8985321

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

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

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

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

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