| BCRJ Code | 0331 |
| Cell Line | PC-9 |
| Species | Homo sapiens |
| Vulgar Name | Human |
| Tissue | Lung |
| Morphology | Heterogeneous mixture of round cells and spindle shaped cells. |
| Disease | Adenocarcinoma |
| Growth Properties | Mixed, Adherent And Suspension |
| Derivation | Derived from a human adenocarcinoma from lung tissue which remains differentiated. |
| Biosafety | 1 |
| Culture Medium | RPMI-1640 medium modified to contain 2 mM L-glutamine, 4500 mg/L glucose and 10% of fetal bovine serum. |
| Subculturing | Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. Remove culture medium with floating cells to a centrifuge tube. Briefly rinse the cell layer with PBS without calcium and magnesium to remove all traces of serum, which 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 10 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. To remove trypsin-EDTA solution, transfer cell suspension to the centrifuge tube with the medium and cells from step #1 and spin at approximately 125 x g for 5 to 10 minutes. Discard supernatant and resuspend cells in fresh growth medium. Add appropriate aliquots of cell suspension to new culture vessels; Seed cells at 2-4 x 10,000 cells/cm2. Place culture vessels in incubator at 37°C. 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 | 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). |
| Depositors | Lidia Moreira Lima - Universidade Federal Do Rio De Janeiro |
| Cellosaurus | CVCL_B260 |
