RTG-P1 (ATCC® CRL-2829)

Organism: Oncorhynchus mykiss, trout, rainbow  / 

Permits and Restrictions

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Organism Oncorhynchus mykiss, trout, rainbow
Product Format frozen
Morphology fibroblast
Culture Properties adherent
Biosafety Level 2 Cells contain SV-40 and CMV viral DNA sequences
Gender male and female mixed
Applications
The vector, pGL3-prMx1-Basic-Neo, expresses the firefly luciferase gene under the control of the promoter for the IFN-induced gene Mx1. The vector contains SV40 viral DNA sequences and the neomycin resistance gene.
This cell line can be used to study the activation of the IFN pathway.
The RTG-2 cell line (ATCC CCL-55) was transfected with a trout Mx1 promoter-luciferase construct and selected to produce a stable cell line, RTG-P1 [PubMed: 15043947].
The RTG-P1 reporter system constitutes an interesting tool to study the induction and regulation of IFN signaling in teleost fish.
Storage Conditions liquid nitrogen vapor phase
Images
Clinical Data
male and female mixed
Comments
The RTG-2 cell line (ATCC CCL-55) was transfected with a trout Mx1 promoter-luciferase construct and selected to produce a stable cell line, RTG-P1 [PubMed: 15043947]. The vector, pGL3-prMx1-Basic-Neo, expresses the firefly luciferase gene under the control of the promoter for the IFN-induced gene Mx1. The vector contains SV40 viral DNA sequences and the neomycin resistance gene. Luciferase expression is stable and highly induced by polyinosinic:polycytidylic acid (poly I:C) in RTG-P1 cells [PubMed: 15043947]. This cell line can be used to study the activation of the IFN pathway. The RTG-P1 reporter system constitutes an interesting tool to study the induction and regulation of IFN signaling in teleost fish.
Complete Growth Medium Leibovitz's L-15 medium with 2 mM L-glutamine supplemented with 0.2 mg/ml G418 and 10% fetal bovine serum.

(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
Protocol:
  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.
  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 6 X 10(3) to 1 X 10(4) viable cells/sq. cm is recommended.
  6. Incubate cultures at 22C.
Interval: Maintain cultures at a cell concentration between 1 X 10(4) and 1 X 10(5) cells/sq. cm
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:4 is recommended
Medium Renewal: Two to three times weekly
Cryopreservation
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 100%
Temperature: 22.0°C
Population Doubling Time approximately 67 hours
Name of Depositor B Collet
References

Collet B, et al. An Mx1 promoter-reporter system to study interferon pathways in rainbow trout. Dev. Comp. Immunol. 28: 793-801, 2004. PubMed: 15043947

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
Other Documentation
References

Collet B, et al. An Mx1 promoter-reporter system to study interferon pathways in rainbow trout. Dev. Comp. Immunol. 28: 793-801, 2004. PubMed: 15043947