Cell Line Name:

B9

Keywords:

Mouse B cell hybridoma, IL-6 assay

Cell
   Line Description:

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.

Species:

Mouse

Tissue of Origin:

Spleen

CellType:

Lymphoblastoid

Growth Mode:

Suspension

Subculture Routine:

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.

Culture Medium:

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.

Country:

NETHERLANDS

References:

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.