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RTG-P1

CRL-2829

Product category
Animal cells
Product type
Reporter-labeled cell
Organism
Oncorhynchus mykiss, trout, rainbow
Morphology
fibroblast
Tissue
Testis; Ovary
Applications
3D cell culture
Assay development
High-throughput screening
Product format
Frozen
Storage conditions
Vapor phase of liquid nitrogen
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Documentation

ATCC determines the biosafety level of a material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to understand the hazards associated with the material per your organization’s policies and procedures as well as any other applicable regulations as enforced by your local or national agencies.

Cells contain cytomegalovirus (CMV) DNA sequences

Cells contain SV40 sequences

ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.

Detailed product information

General

Specific 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.

Characteristics

Growth properties
Adherent
Gender
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.

Handling information

Unpacking and storage instructions
  1. Check all containers for leakage or breakage.
  2. Remove the frozen cells from the dry ice packaging and immediately place the cells at a temperature below ­-130°C, preferably in liquid nitrogen vapor, until ready for use.
Complete 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)

Temperature
22°C
Atmosphere
100% Air
Handling procedure

Handling Procedure for Frozen Cells

          To insure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at -70°C.  Storage at -70°C will result in loss of viability.

SAFETY PRECAUTION: ATCC highly recommends that protective gloves and clothing always be used and a full face mask always be worn when handling frozen vials. It is important to note that some vials leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen.  Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vessel exploding or blowing off its cap with dangerous force creating flying debris.

1.        Thaw the vial by gentle agitation in a 22°C water bath. To reduce the possibility of contamination, keep the O-ring and cap out of the water.  Thawing should be rapid (approximately 2 minutes).

2.        Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions.

3.        Transfer the vial contents to a centrifuge tube containing 9.0 ml complete culture medium. and spin at approximately 125 x g for 5 to 7 minutes.

4.        Resuspend cell pellet with the recommended complete medium (see the specific batch information for the culture recommended dilution ratio) and dispense into a 25 cm2 or a 75 cm2 culture flask.  It is important to avoid excessive alkalinity of the medium during recovery of the cells.  It is suggested that, prior to the addition of the vial contents, the culture vessel containing the complete growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6).

5.       Incubate the culture at 22°C in a suitable incubator.  A free gas exchange with atmospheric air is recommended if using the medium described on this product sheet.

Subculturing procedure
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
Reagents for cryopreservation
Complete growth medium supplemented with 10% (v/v) fetal bovine serum (FBS) and 5% DMSO (ATCC 4-X)

Quality control specifications

Mycoplasma contamination
Not detected
Population doubling time
Approximately 67 hrs

History

Depositors
B Collet

Legal disclaimers

Intended use
This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.
Warranty

The product is provided 'AS IS' and the viability of ATCC® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid.  Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.

Disclaimers

This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. Any proposed commercial use is prohibited without a license from ATCC.

While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.

This product is sent on the condition that the customer is responsible for and assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'AS IS' with no representations or warranties whatsoever except as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, special, incidental, or consequential damages of any kind in connection with or arising out of the customer's use of the product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, ATCC is not liable for damages arising from the misidentification or misrepresentation of such materials.

Please see the material transfer agreement (MTA) for further details regarding the use of this product. The MTA is available at www.atcc.org.

Permits & Restrictions

Import Permit for the State of Hawaii

If shipping to the U.S. state of Hawaii, you must provide either an import permit or documentation stating that an import permit is not required. We cannot ship this item until we receive this documentation. Contact the Hawaii Department of Agriculture (HDOA), Plant Industry Division, Plant Quarantine Branch to determine if an import permit is required.

MORE INFORMATION ABOUT PERMITS AND RESTRICTIONS

Frequently Asked Questions

References

Curated Citations

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

Collet B, et al. A method to measure an indicator of viraemia in Atlantic salmon using a reporter cell line. J Virol Methods 191(2): 113-7, 2013. PubMed: 23602803

Acosta F, et al. Expression of the glycoprotein of viral haemorrhagic septicaemia virus (VHSV) on the surface of the fish cell line RTG-P1 induces type 1 interferon expression in neighbouring cells. Fish Shellfish Immunol 21(3): 272-8, 2006. PubMed: 16580844

For product-related inquiries and issues, contact Technical Service:

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