3B11 cell line细胞株细胞系 BioVector NTCC质粒载体菌种细胞基因保藏中心

3B11 cell line细胞株细胞系 BioVector NTCC质粒载体菌种细胞基因保藏中心
Organism Ictalurus punctatus, channel catfish
Tissue peripheral blood
Cell Type lymphocyte B lymphoblast
Product Format frozen
Morphology lymphoblast
Culture Properties suspension
Biosafety Level 1
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 3B11 cells constitutively express telomerase [PubMed: 11687262].
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 3B11 cells constitutively express telomerase [PubMed: 11687262].
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
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