Code: 0210
Cell Line: RA3-3A1/6.1
Species: Rattus norvegicus (B cell); Mus musculus (myeloma), rat (B cell); mouse (myeloma)

Vulgar Name: Mouse, rat
Tissue: Spleen
Morphology: Lymphoblast
Growth Properties: Suspension
Derivation: Animals were immunized with the mouse cell line RAW 112; spleen cells were fused with the S194/5.XXO.BU.1 myelona cells. This antibody reacts with the B220 antigen on normal B-lymphocytes, a fraction os sIg bone marrow cells and most plasma cells.
Products: immunoglobulin; monoclonal antibody; against the B220 mouse B cell antigen (200000 dalton form of the T200 glycoprotein, CD45R)
Biosafey: 1
Culture Medium: RPMI-1640 medium modified to contain 2 mM L-glutamine, 10 mM HEPES, 1 mM sodium pyruvate, 4500 mg/L glucose, and 1500 mg/L sodium bicarbonate and 10% of fetal bovine serum.
Subculturing: Cultures can be maintained by the addition of fresh medium or replacement of medium. Maintain cell density : procedure: maintain cultures from 1x10e5 to 1x10e6 cells/mL.

Medium Renewal: Every 2 to 3 days

Culture Conditions: Atmosphere: air, 95%; carbon dioxide (CO2), 5% Temperature: 37°C
Cryopreservation: 95% FBS + 5% DMSO (Dimethyl sulfoxide)
Thawing Frozen Cells: SAFETY PRECAUTION: Is highly recommend that protective gloves and clothing always be used and a full face mask always be worn when handling frozen vials. It is important to note that some vials leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vessel exploding or blowing off its cap with dangerous force creating flying debris. 1. Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the Oring and cap out of the water. Thawing should be rapid (approximately 2 minutes). 2. Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions. 3. For cells that are sensitive to DMSO is recommended that the cryoprotective agent be removed immediately. Transfer the vial contents to a centrifuge tube containing 9.0 mL complete culture medium and spin at approximately 125 x g for 5 to 7 minutes. 4.Discard the supernatant and Resuspend cell pellet with the recommended complete medium (see the specific batch information for the culture recommended dilution ratio). 5. Incubate the culture in a appropriate atmosphere and temperature (see "Culture Conditions" for this cell line). NOTE: It is important to avoid excessive alkalinity of the medium during recovery of the cells. It is suggested that, prior to the addition of the vial contents, the culture vessel containing the growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6).
References: Coffman RL, Weissman IL. A monoclonal antibody that recognizes B cells and B cell precursors in mice. J. Exp. Med. 153: 269-279, 1981. PubMed: 6787164 Coffman RL, Weissman IL. B220: a B cell-specific member of the T200 glycoprotein family. Nature 289: 681-683, 1981. PubMed: 6970340 Wong P, Rudensky AY. Phenotype and function of CD4+ T cells in mice lacking invariant chain. J. Immunol. 156: 2133-2142, 1996. PubMed: 8690902 Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC. Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988. Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.
Depositors: Banco de Células do Rio de Janeiro