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1G8

CRL-2756

Product category
Animal cells
Organism
Ictalurus punctatus, channel catfish
Classification
Metazoa, Chordata, Craniata, Actinopterygii, Siluriformes, I
Cell type
B lymphoblast
Morphology
lymphoblast
Tissue
Peripheral blood
Applications
3D cell culture
Immunology
Product format
Frozen
Storage conditions
Vapor phase of liquid nitrogen
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Documentation

Biosafety Icon BSL 1

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.

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
They are reported to secrete moderate levels of IgM in culture [PubMed: 8133033].

Characteristics

Growth properties
Suspension
Derivation
1G8 (ATCC CRL-2756), developed in 1992, and 3B11 (ATCC CRL-2757), developed in 1994, are immortal B cell lines developed by in vitro lipopolysaccharide (LPS) stimulation of peripheral blood from two different normal channel catfish. The cells were subsequently cloned by limiting dilution. The cells are maintained in vitro without restimulation, feeder cells, or exogenous factors. They are reported to secrete moderate levels of IgM in culture [PubMed: 8133033]. The 1G8 cells express p53 mRNA levels only two to three times higher than comparable levels found in freshly isolated peripheral blood [PubMed: 11687262]. The 1G8 cells express the heat shock protein hsp70 gene in significantly higher levels than comparable levels found in freshly isolated peripheral blood [PubMed: 11687262]. The 1G8 cells constitutively express telomerase [PubMed: 9914913].
Karyotype
diploid
Comments
1G8 (ATCC CRL-2756), developed in 1992, and 3B11 (ATCC CRL-2757), developed in 1994, are immortal B cell lines developed by in vitro lipopolysaccharide (LPS) stimulation of peripheral blood from two different normal channel catfish. The cells were subsequently cloned by limiting dilution. The cells are maintained in vitro without restimulation, feeder cells, or exogenous factors. They are reported to secrete moderate levels of IgM in culture [PubMed: 8133033]. The 1G8 cells express p53 mRNA levels only two to three times higher than comparable levels found in freshly isolated peripheral blood [PubMed: 11687262]. The 1G8 cells express the heat shock protein hsp70 gene in significantly higher levels than comparable levels found in freshly isolated peripheral blood [PubMed: 11687262]. The 1G8 cells constitutively express telomerase [PubMed: 9914913].

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
A 1:1 mixture of Leibovitz's L-15 medium with 2 mM L-glutamine, and AIM-V Medium adjusted to contain 1.5 g/L sodium bicarbonate and supplemented with 0.05 mM 2-mercaptoethanol, 87.5%; double distilled water, 10%; heat-inactivated Catfish serum, 2.5%

(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
27°C
Atmosphere
95% Air, 5% CO2
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 27°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 75 cm2 tissue culture flask and dilute with the recommended complete culture medium (see the specific batch information for the recommended dilution ratio).   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 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).

4.        Incubate the culture at 27°C in a suitable incubator.  A 5% CO2 in air atmosphere is recommended if using the medium described on this product sheet.

            If it is desired that the cryoprotective agent be removed immediately, or that a more concentrated cell suspension be obtained, centrifuge the cell suspension at approximately 125 xg for 5 to 10 minutes.  Discard the supernatant and resuspend the cells with fresh growth medium at the dilution ratio recommended in the specific batch  information.

Subculturing procedure
Protocol: Cultures can be maintained by the addition of fresh medium or replacement of medium. Alternatively, cultures can be established by centrifugation with subsequent resuspension at 5 X 10(5) viable cells/ml.
Interval: Maintain cell density between 5 X 10(5) and 5 X 10(6) viable cells/ml.
Medium Renewal: Add fresh medium every 2 to 3 days (depending on cell density)
Reagents for cryopreservation
Complete growth medium supplemented with 5% (v/v) DMSO (ATCC 4-X)

Quality control specifications

Mycoplasma contamination
Not detected

History

Depositors
NW Miller
Year of origin
1992

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

References

Curated Citations

Miller NW, et al. Development and characterization of channel catfish long term B cell lines. J. Immunol. 152: 2180-2189, 1994. PubMed: 8133033

Wilson M, et al. A novel chimeric Ig heavy chain from a teleost fish shares similarities to IgD. Proc. Natl. Acad. Sci. USA 94: 4593-4597, 1997. PubMed: 9114035

Stuge TB, et al. Development and analysis of various clonal alloantigen-dependent cytotoxic cell lines from channel catfish. J. Immunol. 164: 2971-2977, 2000. PubMed: 10706684

Barker K, et al. Immortal and mortal clonal lymphocyte lines from channel catfish: comparison of telomere length, telomerase activity, tumor suppressor and heat shock protein expression. Dev. Comp. Immunol. 26: 45-51, 2002. PubMed: 11687262

Miller N, et al. Functional and molecular characterization of teleost leukocytes. Immunol. Rev. 166: 187-197, 1998. PubMed: 9914913

View All Curated Citations for this Product

Frequently Asked Questions

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Telephone

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