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Restrictions:
Note: These cells are distributed for research purposes only. Dr. Harris releases the line subject to the following: 1.) The Sp2/mIl-6 cells or any cells derived from SP2/mIL-6 or any antibody derived from it must not be sold or used for commercial purposes. Nor can the cells be distributed to third parties for purpose of sale, or producing for sale, cells or their products. 2.) Any proposed commercial use of these cells should first be negotiated with the London Regional Cancer Centre, 790 Commissioners Road, East, London, Ontario. N6A 4L6 Canada. 3.) In all papers reporting any use of these or derived clones a direct reference will be made to the original publication (J. Immunol. Methods 148: 199-207, 1992).
Isolation:
Toronto Ontario, Canada
Antigen Expression:
H-2, d haplotype; Mus musculus, expressed
Comments:
The Sp2/mIL-6 line was established by R. Hawley and T. Hawley of the Sunnybrook Health Centre, Toronto, Canada. It is a mouse fusion cell line derived from Sp2/0 cells that had been transfected with high-titer L6PNL recombinant retrovirus carrying a mouse interleukin-6 (IL-6) cDNA under the transcriptional control of the viral LTR and the neo gene encoding bacterial neomycin phosphotransferase (G418 resistance). Sp2/mIL-6 cells have stably incorporated in their genome approximately six copies of the retroviral vector L6PNL and constitutively secrete large quantities of mouse IL-6 (10,000 units/ml). Sp2/IL-6 cells secrete IL-6 and are an improved myeloma fusion parent in terms of both the relative number of viable B cell hybridomas producing immunoglobulin and the proportion of hybridomas producing antigen-specific monoclonal antibodies.
Propagation:
ATCC complete growth medium: The base medium for this cell line is ATCC Hybri-Care Medium, Catalog No. 46-X. Hybri-Care Medium is supplied as a powder and should be reconstituted in 1 L cell-culture-grade water. To make the complete growth medium, add the following components to the base medium:
fetal bovine serum to a final concentration of 10%
1.5 g/L sodium bicarbonate for use with 5% CO2 in air atmosphere
0.05 mM 2-Mercaptoethanol
Temperature: 37.0°C
Subculturing:
Protocol: Cultures can be maintained by addition or replacement of fresh medium. Start new cultures at 2 X 10(5) cells/ml and subculture before the cell density reaches 2 X 10(6) cells/ml. Medium Renewal: Add fresh medium every 2 to 4 days (depending on cell density)
Related Products:
Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 46-X
References:
23169: Harris JF, et al. Increased frequency of both total and specific monoclonal antibody producing hybridomas using a fusion partner that constitutively expresses recombinant IL-6. J. Immunol. Methods 148: 199-207, 1992. PubMed: 1373425
