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MDCK.2

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編號:B181962

品牌:Mingzhoubio

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RNA

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產品名稱

MDCK.2

商品貨號

B181962

Organism

Canis familiaris, dog

Tissue

kidney; distal tubule

培養(yǎng)基

Product Format

frozen

Morphology

epithelial-like

Culture Properties

adherent

Biosafety Level

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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