Cell Line Name:
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B9
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Keywords:
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Mouse B cell hybridoma, IL-6 assay
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Cell
Line Description:
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B9 is a mouse B cell hybridoma line that requires Interleukin 6 (IL-6)
for survival and proliferation. IL-6 is a pluripotent cytokine with multiple
effects on many different cell types. It is produced by a variety of cells in
response to various stimuli. The B9 hybridoma allows for a simple sensitive
assay for human, rat, rabbit, pig and mouse IL-6 based on the IL-6 dependent
proliferation of this murine B cell line. Details of the B9 hybridoma growth
factor assay can be found in Current protocols in Immunology 1996 Cytokines
and Their Cellular receptors 6.6.1-6.6.5 contributed by Nordan RP, Richards
CD and Gauldie J. John Wiley & Sons, Inc.
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Species:
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Mouse
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Tissue of Origin:
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Spleen
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CellType:
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Lymphoblastoid
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Growth Mode:
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Suspension
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Subculture Routine:
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For B9 cell maintenance cells should be seeded at 0.5 x 105cells/ml in B9 culture medium. The culture will be in log phase growth
between days 3-5. When the culture reaches 1 x 106cells/ml (4-5 days) the culture must be sub cultured as above again
seeding at 0.5 x 105cells/ml.; 5% CO2;37°C. It maybe
necessary to determine the activity of the IL-6 or IL-6 containing
supernatant first. The lowest concentration at which maximal growth can be
obtained should be used. A fresh vial of cells is required when the
background response from control cultures begins to rise. It is important
that the culture conditions are adhered to avoid the B9 cells becoming IL-6
independent.
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Culture Medium:
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IDMEM or RPMI 1640 + 2mM Glutamine + 50µM 2-Mercaptoethanol (2ME) +
50pg/ml recombinant human IL-6 (Sigma SRP3096, diluted and stored in PBS
containing 1% BSA) + 5% FBS. IL-6 needs to be added fresh at each media
change.
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Country:
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NETHERLANDS
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References:
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Aarden LA, De Groot ER, Schaap OL, Lansdorp PM. 1987 Production of
hybridoma growth factor by human monocytes. Eur J Immunol. 17(10):1411-6.PMID: 3500054.
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