The cells are free from the classical swine fever viruses and porcine circoviruses as described in 9 CFR 113.46, 9 CFR 113.47 and 9 CFR 113.55.xa0

The SK-RST cells exhibited cytopathology after being infected with two serotypes of vesicular stomatitis virus, Newcastle disease virus, porcine parvovirus, pseudorabies virus, reovirus, transmissible gastroenteritis virus, bovine virus diarrhea, bovine adenovirus, bovine parainfluenza 3 virus, infectious bovine rhinotracheitis virus, and with representatives of the A, O, C, SAT1, SAT2, SAT3 and Asia serotypes of foot-and-mouth disease virus.

Classical swine fever virus (CSF)-infected SK-RST cells produced CSF antigen that was detectable using a CSF-antigen specific monoclonal antibody in an avidin-biotin complex immunohistochemistry assay.xa0

The SK-RST cells failed to exhibit cytopathology when exposed to porcine adenovirus, bluetongue virus, bovine respiratory syncytial virus, and bovine parvovirus.xa0

1. Remove and discard culture medium.
2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
3. 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.
4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
5. Add appropriate aliquots of the cell suspension to new culture vessels.
   An inoculum of 5 x 10³xa0to 7 x 10³xa0viable cells/cm² is recommended.
6. Incubate cultures at 37°C.

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.