0156 - M3/84.6.34

BCRJ Code 0156
Cell Line M3/84.6.34
Species Rattus norvegicus (B cell); Mus musculus (myeloma), rat (B cell); mouse (myeloma)
Vulgar Name Rat/Mouse
Cell Type Hybridoma: B Lymphocyte
Morphology Lymphoblast
Growth Properties Suspension
Applications Mac-2 and Mac-3 are present on 69% of macrophages and 0% to 2% of thymocytes.
Products immunoglobulin; monoclonal antibody; against Mac-3 (mouse macrophage antigen, 110000 dalton glycoprotein)
Biosafety 1
Addtional Info Animals were immunized with detergent solubilized mouse(C57BL/6) macrophage membrane which had been depleted of previously identified antigens with monoclonal immunoadsorbants. Like Mac-2, the Mac-3 antigen is not expressed on bone marrow cells (Mac-1 is expressed on bone marrow cells). Also like Mac-2, Mac-3 appears to be expressed on their monocytic line of differentiation at a stage after divergence from the granulocytic series. Mac-2 and Mac-3 are present on 69% of macrophages and 0% to 2% of thymocytes. Expression of Mac-3 is increased during the differentiation from monocyte to activated peritoneal macrophage.
Culture Medium RPMI-1640 medium modified to contain 2 mM L-glutamine, 1 mM sodium pyruvate, 4500 mg/L glucose and 10% of fetal bovine serum.
Subculturing Cultures can be maintained by the addition of fresh medium or replacement of medium. Adherent cells can be dislodged by scraping and cultures established by centrifugation with subsequent resuspension at 1 to 2 X 10(5) viable cells/ml.
Subculturing 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 Springer TA. Monoclonal antibody analysis of complex biological systems. Combination of cell hybridization and immunoadsorbents in a novel cascade procedure and its application to the macrophage cell surface. J. Biol. Chem. 256: 3833-3839, 1981. PubMed: 7217058
Depositors Banco de Células do Rio de Janeiro