SOB-15 (ATCC® CRL-2301)

Organism: Oncorhynchus mykiss, trout, rainbow  /  Cell Type: epithelial  /  Tissue: liver, epithelial  / 

Permits and Restrictions

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Organism Oncorhynchus mykiss, trout, rainbow
Tissue liver, epithelial
Cell Type epithelial
Product Format frozen
Morphology spindle-like with polygonal foci
Culture Properties adherent
Biosafety Level 1
Strain Shasta
Storage Conditions liquid nitrogen vapor phase
Comments
The cells retain characteristics of normal fish liver
Complete Growth Medium Minium essential Medium Eagle with 2 mM L-glutamine and Earle's BSS adjusted to contain 1.5 g/L sodium bicarbonate, 0.1 mM non-essential amino acids, and 1.0 mM sodium pyruvate and supplemented with 2 mM L-glutamine, 1.32 g/L sodium chloride, 0.1 mM non-essential amino acids and 10% heat-inactivated fetal bovine serum.
Subculturing Volumes used in this protocol are for 75 cm2 flask; 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 0.25% (w/v) Trypsin-0.53mM EDTA solution to remove all traces of serum which 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 17°C.

Subcultivation Ratio: 1:2 to 1:3
Medium Renewal: Every 2 to 3 days

Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a Manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.

Cryopreservation

Complete growth medium described above supplemented with 5% (v/v) DMSO.  Cell culture tested DMSO is available as ATCC Catalog No. 4-X.

Culture Conditions
Temperature: 17°C
Atmosphere: Air, 95%; Carbon dioxide (CO2), 5%
Name of Depositor GK Ostrander, JB Blair
References

Ostrander GK, et al. Long-term primary culture epithelia cells from rainbow trout (Oncorhynchus mykiss) liver. In Vitro Cell. Dev. Biol. Anim. 31: 367-378, 1995. PubMed: 7543343

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
References

Ostrander GK, et al. Long-term primary culture epithelia cells from rainbow trout (Oncorhynchus mykiss) liver. In Vitro Cell. Dev. Biol. Anim. 31: 367-378, 1995. PubMed: 7543343

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.