Disease: plasmacytoma; myeloma Strain: BALB/c Cell Type: B lymphocyte;
Cellular Products:
immunoglobulin (not secreted)
Permits/Forms:
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Antigen Expression:
H-2d
Comments:
This is a non-secreting clone of P3X63Ag8 (ATCC TIB-9).
Kappa chains are synthesized but not secreted.
The cells are resistant to 0.1 mM 8-azaguanine and do not grow in HAT medium.
The cells have been reported to be cholesterol auxotrophs due to a deficiency in 3-ketosteroid reductase activity.
Tested and found negative for ectromelia virus (mousepox).
Propagation:
ATCC complete growth medium: The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%. Temperature: 37.0°C Atmosphere: air, 95%; carbon dioxide (CO2), 5%
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 2 X 10(5) viable cells/ml. Maintain cell density between 1 X 10(5) and 1 X 10(6) viable cells/ml. Do not allow the cell density to exceed 1 X 10(6) cells/ml. Medium Renewal: Every 2 to 3 days
Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2002
recommended serum:ATCC 30-2020
parental cell line:ATCC TIB-9
References:
956: Barnstable CJ, et al. Production of monoclonal antibodies to group A erythrocytes, HLA and other human cell surface antigens -- new tools for genetic analysis. Cell 14: 9-20, 1978. PubMed: 667938 1115: Kohler G, et al. Fusion between immunoglobulin-secreting and nonsecreting myeloma cell lines. Eur. J. Immunol. 6: 292-295, 1976. PubMed: 825374 22154: Horibata K, Harris AW. Mouse myelomas and lymphomas in culture. Exp. Cell Res. 60: 61-77, 1970. PubMed: 5439579 22347: Kohler G, Milstein C. Derivation of specific antibody-producing tissue culture and tumor lines by cell fusion. Eur. J. Immunol. 6: 511-519, 1976. PubMed: 825377 23366: Sato JD, et al. Effects of proximate cholesterol precursors and steroid hormones on mouse myeloma growth in serum-free medium. In Vitro Cell. Dev. Biol. 24: 1223-1228, 1988. PubMed: 3209588 26333: Ramasamy R, et al. Possible role for the Fc receptor on B lymphocytes. Nature 249: 573-574, 1974. PubMed: 4545851 26344: Cowan NJ, et al. Intracellular immunoglobulin chain synthesis in non-secreting variants of a mouse myeloma: detection of inactive light-chain messenger RNA. J. Mol. Biol. 90: 691-701, 1974. PubMed: 4449137 33092: Condie R, et al. Cannabinoid inhibition of adenylate cyclase-mediated signal transduction and interleukin 2 (IL-2) expression in the murine T-cell line, EL4.IL-2. J. Biol. Chem. 271: 13175-13183, 1996. PubMed: 8662742
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