| BCRJ Code | 0016 |
| Cell Line | 3H5-1 |
| Species | Mus musculus (B cell); Mus musculus (myeloma), mouse (B cell); mouse (myeloma) |
| Vulgar Name | Mouse |
| Cell Type | Hybridoma: B Lymphocyte |
| Morphology | Lymphoblast |
| Disease | Dengue |
| Growth Properties | Suspension |
| Derivation | Spleen cells were fused with P3X63Ag8 myeloma cells. |
| Tumor Formation: | YES |
| Products | immunoglobulin; monoclonal antibody; against a type specific determinant on Dengue virus-2) |
| Biosafety | 1 |
| Addtional Info | Animals were immunized with dengue virus type 2 antigens from infected mouse brains (the prototype strain New Guinea C was used). The antibody is type specific. Spleen cells were fused with P3X63Ag8 myeloma cells. Antibodies should be prepared as ascites |
| Culture Medium | Hybri-Care Medium from ATCC (Catalog No. 46-X), 1.5 g/L sodium bicarbonate 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 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.
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 | 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 | Ada M.B.Alves |
| Cellosaurus | CVCL_D292 |
