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African green monkey kidney cell line suitable for transfection by SV40 vectors.

Volumes used in this protocol are for 75 cm² flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.

1. Remove and discard culture medium. 
2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin-0.53mM EDTA solution to remove all traces of serum which 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.
6. Incubate cultures at 37°C.

Hopps, H. E., et al. Biologic characteristics of the continuous kidney cell line derived from the African green monkey. J. Immunol. 91: 416-424, 1963. PubMed: 14071033

American Public Health Association. Compendium of methods for the microbiological examination of foods. 3rd ed.Washington, DC: American Public Health Association; 1992.

Berson JF, et al. A seven-transmembrane domain receptor involved in fusion and entry of T-cell-tropic human immunodeficiency virus tyep 1 strains. J. Virol. 70: 6288-6295, 1996. PubMed: 8709256

Jelachich ML, Lipton HL. Theiler's murine encephalomyelitis virus kills restrictive but not permissive cells by apoptosis. J. Virol. 70: 6856-6861, 1996. PubMed: 8794327

Schmidt NJ, et al. The sensitivity of grivet monkey kidney cell line BS-C-1 for propagation and isolation of certain human viruses. Am. J. Public Health 54: 1522-1530, 1964.