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-0.53mMxa0 EDTA solution to remove all traces of serum whichxa0 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 cellxa0xa0 layer is dispersed (usually within 5 to 15 minutes).xa0Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach.xa0 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.xa0
5. Add appropriate aliquots of the cell suspension to new culture vessels.
6. Incubate cultures at 37°C.

Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:3 is recommended

Medium Renewal: 2 to 3 times per week

Hoying J. Selection and histochemical identification of epithelial-like cells from guinea pig lung M.Sc. thesis, Wright State Univ., 1975

Nucleotide (GenBank) : AF131949 Cavia porcellus tissue factor mRNA, complete cds.