1G8 (ATCC® CRL-2756)

Organism: Ictalurus punctatus, channel catfish  /  Cell Type: lymphocyte B lymphoblast  / 

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Organism Ictalurus punctatus, channel catfish
Cell Type lymphocyte B lymphoblast
Product Format frozen
Morphology lymphoblast
Culture Properties suspension
Biosafety Level 1
Applications
They are reported to secrete moderate levels of IgM in culture [PubMed: 8133033].
Storage Conditions liquid nitrogen vapor phase
Karyotype diploid
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].
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].
Complete Growth 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)


Subculturing
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)
Cryopreservation
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 27.0°C
Name of Depositor NW Miller
Year of Origin 1992
References

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

Shen L, et al. Channel catfish cytotoxic cells: a mini-review. Dev. Comp. Immunol. 26: 141-149, 2002. PubMed: 11696379

Miller NW, et al. Development of leukocyte cell lines from the channel catfish (Ictalurus punctatus). J. Tissue Culture Methods 16: 117-123, 1994.

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

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

Shen L, et al. Channel catfish cytotoxic cells: a mini-review. Dev. Comp. Immunol. 26: 141-149, 2002. PubMed: 11696379

Miller NW, et al. Development of leukocyte cell lines from the channel catfish (Ictalurus punctatus). J. Tissue Culture Methods 16: 117-123, 1994.