| BCRJ Code | 0114 |
| Cell Line | IB3-1 [JHU-52] |
| Species | Homo sapiens |
| Vulgar Name | Human |
| Tissue | Lung |
| Morphology | Epithelial |
| Disease | Fibrosis |
| Growth Properties | Adherent |
| Sex | Male |
| Derivation | IB3-1 is an immortalized cell line created in 1992 from a primary culture of bronchial epithelia cells isolated from a patient with cystic fibrosis. The culture was transformed with a hybrid virus, adeno-12-SV40 [PubMed: 1849726] |
| Applications | Transient Transfection |
| Virus Succeptility: | CELLS CONTAIN SV40 AND ADENOVIRUS 12 DNA VIRAL SEQUENCES |
| Biosafety | 2 |
| Culture Medium | LHC-8 Basal Medium (Invitrogen catalog #12679-015 ), 95%; fetal bovine serum, 5% |
| Subculturing | Remove and discard culture medium. Briefly rinse the cell layer with PBS without calcium and magnesium to remove all traces of serum that contains trypsin inhibitor. Add 2.0 to 3.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes). Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting. Add appropriate aliquots of the cell suspension to new culture vessels. Incubate cultures at 37°C. Is recommended Maintain cultures at a cell concentration between 4 X 10(3) and 4 X 10(4) cells/cm2 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 | 2 to 3 times per 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.
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 | 39291: Flotte TR, et al. Gene expression from adeno-associated virus vectors in airway epithelial cells. Am. J. Respir. Cell Mol. Biol. 7: 349-356, 1992. PubMed: 1325813 70684: Afione SR, et al. Expression of the cystic fibrosis transmembrane conductance |
| Depositors | Luiz Soares; Grupo Boticario. |
| Cellosaurus | CVCL_0338 |
