male
52 years

The cells are positive for keratin by immunoperoxidase staining.

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. Add appropriate aliquots of the cell suspension to new culture vessels.
5. Incubate cultures at 37°C.

NOTE: Cells attach in clusters. Cells will pile and the culture does not get 100% confluent.

Moore GE, Sandberg AA. Studies of a human tumor cell line with a diploid karyotype. Cancer 17: 170-175, 1964. PubMed: 14123677

Moorhead PS. Human tumor cell line with a quasi-diploid karyotype (RPMI 2650). Exp. Cell Res. 39: 190-196, 1965. PubMed: 5831238