RTgill-W1 (ATCC® CRL-2523)

Organism: Oncorhynchus mykiss, trout, rainbow  /  Tissue: gill  /  Disease: normal

Permits and Restrictions

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Organism Oncorhynchus mykiss, trout, rainbow
Tissue gill
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 1
Disease normal
Strain Walbaum
Applications

The cell line may be used for in vitro toxicology, fish virology and fish biochemistry.


Karyotype heteroploid
Derivation
RTgill-W1 was established from a 15 month old primary culture of apparently normal rainbow trout gill fragments.
Virus Susceptibility Infectious salmon anemia virus , Infectious salmon anemia virus
Comments

The cell line supports the replication of infectious salmon anemia (ISA) virus but CPE is not observed.

RTgill-W1 is a modest generator of eicosanoids in that only low levels of thromboxane B2 and prostaglandin E2 are detected while no lipoxygenase products are observed.

Prior to deposit at ATCC, the line was found to be contaminated with mycoplasma, and was cured by treatment with BM-Cycline and mycoplasma removal agent (MRA).
Complete Growth Medium The base medium for this cell line is ATCC-formulated Leibovitz's L-15 Medium, Catalog No. 30-2008. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.

(Note: The L-15 medium formulation was devised for use in a free gas exchange with atmospheric air. A CO2 and air mixture is detrimental to cells when using this medium for cultivation)


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-0.53 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 18-20°C without CO2 in the atmosphere.

Subculture Ratio: 1:3 to 1:4
Medium Renewal: Twice a week.
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
culture medium 95%; DMSO, 5%
Culture Conditions
Temperature: 19°C; (Max. 20°C, Min. 18°C)
Atmosphere: air, 100%
Name of Depositor NC Bols
References

Bols NC, et al. Development of a cell line from primary cultures of rainbow trout, Oncorhynchus mykiss (Walbaum), gills. J. Fish Dis. 17: 601-611, 1994.

Falk K, et al. Characterization of infectious salmon anemia virus, an orthomyxo-like virus isolated from Atlantic salmon (Salmo salar L.). J. Virol. 71: 9016-9023, 1997. PubMed: 9371558

Schirmer K, et al. Ability of 16 priority PAHs to be photocytotoxic to a cell line from the rainbow trout gill. Toxicology 127: 143-155, 1998. PubMed: 9699801

Holland JW, et al. The eicosanoid generating capacity of isolated cell populations from the gills of the rainbow trout, Oncorhynchus mykiss. Comp. Biochem. Physiol. C. Pharmacol. Toxicol. Endocrinol. 122: 297-306, 1999. PubMed: 10336089

Schirmer K, et al. Ability of 16 priority PAHs to be directly cytotoxic to a cell line from the rainbow trout gill. Toxicology 127: 129-141, 1998. PubMed: 9699800

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

Bols NC, et al. Development of a cell line from primary cultures of rainbow trout, Oncorhynchus mykiss (Walbaum), gills. J. Fish Dis. 17: 601-611, 1994.

Falk K, et al. Characterization of infectious salmon anemia virus, an orthomyxo-like virus isolated from Atlantic salmon (Salmo salar L.). J. Virol. 71: 9016-9023, 1997. PubMed: 9371558

Schirmer K, et al. Ability of 16 priority PAHs to be photocytotoxic to a cell line from the rainbow trout gill. Toxicology 127: 143-155, 1998. PubMed: 9699801

Holland JW, et al. The eicosanoid generating capacity of isolated cell populations from the gills of the rainbow trout, Oncorhynchus mykiss. Comp. Biochem. Physiol. C. Pharmacol. Toxicol. Endocrinol. 122: 297-306, 1999. PubMed: 10336089

Schirmer K, et al. Ability of 16 priority PAHs to be directly cytotoxic to a cell line from the rainbow trout gill. Toxicology 127: 129-141, 1998. PubMed: 9699800