Catálogo de Células

0208 - R24

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BCRJ Code 0208
Cell Line R24
Species Mus musculus (B cell); Mus musculus (myeloma), mouse (B cell); mouse (myeloma)
Vulgar Name Rat
Cell Type Hybridoma: B Lymphocyte
Morphology Lymphoblast
Growth Properties Suspension
Derivation Spleen cells were fused with NS-1 myeloma cells.
Applications The antibody reacts with the M-18 antigen system
Products immunoglobulin; monoclonal antibody; against a human melanoma cell line (M-18 antigen system)
Biosafety 1
Addtional Info Animals were immunized with the SK-MEL-28 human melanoma cell line. Spleen cells were fused with NS-1 myeloma cells. The antibody reacts with the M-18 antigen system.
Culture Medium RPMI-1640 medium modified to contain 2 mM L-glutamine, 4500 mg/L glucose and 10% of fetal bovine serum.
Subculturing Cultures can be maintained by addition or replacement of fresh medium. Start cultures at 2 X 10 exp5 cells/ml and maintain between 1 X 10 exp5 and 1 X 10 exp6 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 J.Exp.Med. 155:1133-1147, 1982 ibid., 156:1755-1766, 1982 Behring Inst. Mitt. 74:14-18, 1984 Cancer REs. 190-194, 1984 Eur. J. Clin. Onc. 21:907-912, 1985 Proc. Natl. Acad. Sci. USA 82:1242-1246, 1985
Depositors Banco de Células do Rio de Janeiro
Cellosaurus CVCL_D295
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