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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.
6. Incubate cultures at 37°C.

Rhim JS, et al. Non-producer human cells induced by murine sarcoma virus. Int. J. Cancer 15: 23-29, 1975. PubMed: 165148

McAllister RM, et al. Cultivation in vitro of cells derived from a human osteosarcoma. Cancer 27: 397-402, 1971. PubMed: 5100401

Rhim JS, et al. Characterization of non-producer human cells induced by Kirsten sarcoma virus. Int. J. Cancer 16: 840-849, 1975. PubMed: 171229

Rhim JS. Characterization of sarcoma-positive, leukemia-negative (S+L-) human cells induced by the feline leukemia virus pseudotype of Moloney sarcoma virus. Proc. Soc. Exp. Biol. Med. 167: 597-606, 1981. PubMed: 6269117

Rhim JS, et al. Differential susceptibility of human cells to transformation by murine and avian sarcoma viruses. Proc. Soc. Exp. Biol. Med. 170: 350-358, 1982. PubMed: 6283561

. . Nat. New Biol. 230: 279-282, 1971.

Yee A, et al. Biochemical characterization of the human cyclin-dependent protein kinase activating kinase. J. Biol. Chem. 271: 471-477, 1996. PubMed: 8550604