0268 - D1-4G2-4-15

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BCRJ Code 0268
Cell Line D1-4G2-4-15
Species Mus musculus (B cell); Mus musculus (myeloma), mouse (B cell); mouse (myeloma)
Vulgar Name Mouse
Tissue Spleen
Cell Type B Lymphocyte; Hybridoma
Morphology Lymphoblast-Like
Growth Properties Suspension
Derivation Animals were immunized with dengue virus type 2 antigens. Spleen cells were fused with P3X63Ag8 myeloma cells.
Products immunoglobulin; antibody against flavivirus group antigen
Biosafety 1
Addtional Info The antibody reacts with flavivirus group specific antigens.
Culture Medium Hybri-Care Medium from ATCC (Catalog No. 46-X) and fetal bovine serum to a final concentration of 10%.
Subculturing Cultures can be maintained by addition of fresh medium. Alternatively, cultures can be established by centrifugation with subsequent resuspension at 10e5 viable cells/mL. Maintain cultures at a cell concentration between 2 x 10e5 and 1 x 10e6cells/mL.
Subculturing Medium Renewal 2 to 3 times a week
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 922: Henchal EA, et al. Dengue virus-specific and Flavivirus group determinants identified with monoclonal antibodies by indirect immunofluorescence. Am. J. Trop. Med. Hyg. 31: 830-836, 1982. PubMed: 6285749
Depositors Danielle da Glória D Souza, Universidade Federal de Minas Gerais
ATCC HB-112
Cellosaurus CVCL_J890