| BCRJ Code | 0340 |
| Cell Line | IB-RS-2 C-17 |
| Species | Sus scrofa |
| Vulgar Name | Pig |
| Tissue | Kidney |
| Cell Type | Epithelial |
| Morphology | Epithelial |
| Disease | Normal |
| Growth Properties | Adherent |
| Virus Succeptility: | Foot and mouth disease virus; FMDV |
| Biosafety | 1 |
| Addtional Info | IB-RS-2 cell line was spontaneously established from primary cultures of kidney cortex of a three-month old pig. It is suceptible to most foot-and-mouth-desease virus (FMDV) types and it is useful for studies of this desease. IB-RS-2 C17 is a clone of IB-RS-2 cell line. |
| Culture Medium | Dulbecco's Modified Eagle's Medium (DMEM) modified to contain 2 mM L-glutamine, 4500 mg/L glucose and fetal bovine serum to a final concentration of 10%. |
| Subculturing | Volumes are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes. 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. 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 |
| Subculturing Subcultivation Ratio | 1:3 to 1:6 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 | Arq. Inst. Biol. Sao Paulo 31: 63-78, 1964; Arq. Inst. Biol. Sao Paulo 31: 155-166, 1964 |
| Depositors | Centro de Pesquisa de Febre Aftosa, through Dr. Amilcar Tanuri, Universidade Federal do Rio de Janeiro |
| Cellosaurus | CVCL_U235 |
