Organism

Fathead minnow

Tissue

skin

Cell Type

epithelial

Product Format

frozen

Morphology

epithelial

Culture Properties

adherent

Biosafety Level

1

Applications

The EPC line has a
   broad sensitivity for fish viruses. It is used internationally for isolation,
   propagation and diagnostic assays for fish viruses.

Virus Susceptibility

Infectious
   pancreatic necrosis virus

   Infectious hematopoietic necrosis virus

   Spring Viremia of Carp Virus

   Viral hemorrhagic septicemia virus

   Epizootic hematopoietic necrosis virus

   Largemouth bass virus

Complete Growth Medium

The base medium for
   this cell line is NTCC-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 cm2 flasks; proportionally reduce or increase amount
   of dissociation medium for culture vessels of other sizes.

1.Remove and discard culture medium.

2.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.

3.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 minutes).

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. An
   inoculum of 5 x 103 to 7 x 103 viable cells/cm2 is
   recommended.

6.
Incubate
   cultures at 25°C. We recommend that you maintain cultures at a cell
   concentration between 1 x 105 and 2 x 105cells/cm2.

   Subcultivation Ratio: 1:4 to 1:6

Cryopreservation

Freeze medium: culture
   medium, 95%; DMSO,5%

Storage temperature: liquid
   nitrogen vapor phase

Culture Conditions

Atmosphere: air,
   95%; carbon dioxide (CO2), 5%

Temperature: 25°C

Max Temperature: 30°C

Min Temperature: 20°C

Population Doubling Time

about 36 hours

References

Fijan N, et al. Some
   properties of the Epithelioma Papulosum Cyprini (EPC) cell line from carp
   Cyprinus carpio. Ann. Virol. (Inst. Pasteur) 134 E: 207-220, 1983.

Fryer JL, Lannan CN.
   Three decades of fish cell culture: A current listing of cell lines derived
   from fishes. J. Tissue Culture Methods 16: 87-94, 1994.