Catálogo de Células

0012 - 331.12

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BCRJ Code 0012
Cell Line 331.12
Species Rattus norvegicus (B cell); Mus musculus (myeloma), rat (B cell); mouse (myeloma)
Vulgar Name Rat/Mouse
Tissue Blood
Cell Type Hybridoma: B Lymphocyte
Morphology Lymphoblast
Disease Lymphoma
Growth Properties Suspension
Derivation Animals were immunized with W279 B lymphoma cells.
Products immunoglobulin; monoclonal antibody; against mouse IgM (mu heavy chain)
Biosafety 1
Culture Medium Dulbecco's Modified Eagle's Medium (DMEM) modified to contain 4 mM L-glutamine, 4500 mg/L glucose, 1 mM sodium pyruvate with 10% of fetal bovine serum.
Subculturing Cultures can be maintained by addition of fresh medium. Alternatively, cultures can be established by centrifugation with subsequent resuspension at 1 x 10e5 viable cells/mL. Maintain cultures at a cell concentration between 1 x 10e5 and 1 x 10e6 cells/mL. NOTE: Do not allow the cell concentration to exceed 1 x 10e6 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: It is strongly recommended to always wear protective gloves, clothing, and a full-face mask when handling frozen vials. Some vials may leak when submerged in liquid nitrogen, allowing nitrogen to slowly enter the vial. Upon thawing, the conversion of liquid nitrogen back to its gas phase may cause the vial to explode or eject its cap with significant force, creating flying debris.
  1. Thaw the vial by gently agitating it in a 37°C water bath. To minimize contamination, keep the O-ring and cap out of the water. Thawing should be rapid (approximately 2 minutes).
  2. Remove the vial from the water bath as soon as its contents are thawed and decontaminate it by dipping in or spraying with 70% ethanol. From this point, all operations must be performed under strict aseptic conditions.
  3. For cells sensitive to DMSO, it is recommended to remove the cryoprotective agent immediately. Transfer the vial contents to a centrifuge tube containing 9.0 mL of complete culture medium and centrifuge at approximately 125 × g for 5 to 7 minutes.
  4. Discard the supernatant and resuspend the cell pellet in the recommended complete medium (see specific batch information for the appropriate dilution ratio).
  5. Incubate the culture under appropriate atmospheric and temperature conditions (see "Culture Conditions" for this cell line).

NOTE: It is important to avoid excessive alkalinity of the medium during cell recovery. To minimize this risk, it is recommended to place the culture vessel containing the growth medium in the incubator for at least 15 minutes before adding the vial contents. This allows the medium to stabilize at its normal pH (7.0 to 7.6).
References Kincade PW, et al. Monoclonal rat antibodies to murine IgM determinants. J. Immunol. Methods 42: 17-26, 1981. PubMed: 6165776
Depositors Alberto Nobrega, Universidade Federal do Rio de Janeiro
Cellosaurus CVCL_9158
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