| BCRJ Code | 0347 |
| Cell Line | JAWSII |
| Species | Mus musculus |
| Vulgar Name | Mouse;C57Bl/6 |
| Tissue | Bone Marrow |
| Cell Type | Immature Dendritic Cell; Monocyte |
| Morphology | Monocyte |
| Growth Properties | Mixed, Adherent And Suspension |
| Biosafety | 1 |
| Culture Medium | Alpha minimum essential medium with ribonucleosides, deoxyribonucleosides, 2 mM L-glutamine, 5 ng/ml murine GM-CSF and 10% of fetal bovine serum. |
| Subculturing | Cultures can be maintained by transferring floating cells to a centrifuge tube. Attached cells may be subcultured using 0.25% trypsin-0.03% EDTA. Pool cells and centrifuge the cell suspension at 1000 rpm for 10 minutes, resuspend the pellet in fresh medium, aspirate and dispense into new flasks. Note: This cell line grows very slowly. NOTE: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 12 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 6th edition, published by Alan R. Liss, N.Y., 2010. |
| Subculturing Medium Renewal | Once a week |
| Subculturing Subcultivation Ratio | 1:2 is recommended |
| 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 | MacKay VL, Moore EE. Immortalized dendritic cells. US Patent 5,648,219 dated Jul 15 1997 Moore EE. Preparation of immortalized cells. US Patent 5,830,682 dated Nov 3 1998 |
| Depositors | Luiz Carlos Rodrigues Junior - Centro Universitário Fransciscano |
| Cellosaurus | CVCL_3727 |
