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1G8

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編號:B238827

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產品名稱	1G8
商品貨號	B238827
Organism	Ictalurus punctatus, channel catfish
Tissue	peripheral blood
Cell Type	lymphocyte B lymphoblast
Product Format	frozen
Morphology	lymphoblast
Culture Properties	suspension
Biosafety Level	1 Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
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.

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