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UK Health Security Agency

B9

B9

Catalogue No.

12121201

Cell Line Name

B9

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.

Characteristics

Tissue of Origin

Spleen

Karyotype

Not specified

Applications

Bioassay for IL-6

Disease

None Stated

Culture Conditions

Cell Type

Lymphoblastoid

Subculture Routine

On resuscitation, cells should be seeded at 5e+05 cells/ml. Once established, cultures should be maintained between 5e+04 and 1e+06 cells/ml.

B9 cells should not be left for a period longer than 4 days without fresh IL-6. IL-6 should be added at each passage or should be replenished every 2-4 days.

It may be 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.

Recommended Growth Conditions: 5% CO2; 37°C.

Culture Medium

IDMEM or RPMI 1640 + 2mM Glutamine + 50µM 2-Mercaptoethanol (2ME) + 50pg/ml recombinant human IL-6 + 5% FBS.

IL-6 (Sigma SRP3096 or equivalent, diluted and stored in PBS containing 1% BSA) must refrigerated separately from the media and should be added at each passage.

Growth Mode

Suspension

Additional Info

Depositor

Professor L A Aarden, Department Autoimmune Diseases, Central Laboratory of the Netherlands Red Cross Blood Transfusion Service, Amsterdam, The Netherlands

Country of Origin

Netherlands

Hazard Group (ACDP)

Hazard Group (ACDP) 2

Applications

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.

Bibliography

Helle M, Boeije L, Aarden LA (1988) Functional discrimination between interleukin 6 and interleukin 1. Eur J Immunol. 18(10):1535-40 PMID: 3263920.

Available Formats

  • Frozen
  • DNA-5µg (100ng/µl)

If use of this culture results in a scientific publication, it should be cited in the publication as: B9 (ECACC 12121201).

Unless specified otherwise, at ECACC we routinely handle all of our cell lines at containment level 2 in accordance with the ACDP guidelines (Advisory Committee on Dangerous Pathogens) (UK). All cell cultures have the potential to carry as yet unidentified adventitious agents. It is the responsibility of the end user to ensure that their facilities comply with biosafety regulations for their own country. ACDP Guidance: Biological agents: Managing the risks in laboratories and healthcare premises.

The Culture Collections represent deposits of cultures from world-wide sources. While every effort is made to ensure details distributed by Culture Collections are accurate, Culture Collections cannot be held responsible for any inaccuracies in the data supplied. References where quoted are mainly attributed to the establishment of the cell culture and not for any specific property of the cell line, therefore further references should be obtained regarding cell culture characteristics. Passage numbers where given act only as a guide and Culture Collections does not guarantee the passage number stated will be the passage number received by the customer.

Cultures supplied by Culture Collections are for research purposes only. Enquiries regarding the commercial use of a cell line are referred to the depositor of the cell line. Some cell lines have additional special release conditions such as the requirement for a material transfer agreement to be completed by the potential recipient prior to the supply of the cell line. Please view the Terms & Conditions of Supply for more information.