Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

The FHM (Fat Head Minnow) line of epithelial cells was derived from tissue posterior to the anus, exclusive of the caudal fin, of normal adult minnows of both sexes by M. Gravell and R.G. Malsberger, in April, 1962.

poliovirus 1

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- 0.53 mM 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 andxa0 allow the flask to sit at room temperature. Observe cells under an inverted microscope until cell 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
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.xa0
6. Incubate cultures at 34°C.

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

Medium Renewal: At the time of subcultivation (this will vary depending upon the temperature of incubation)

Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO

Storage temperature: liquid nitrogen vapor phase

Atmosphere: air, 100%

Temperature: 34°C; (Max. 34°C, Min. 28°C)