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.
2. Briefly rinse the cell layer with 5.0 ml of Ca++/Mg++ free Dulbecco's phosphate-buffered saline (DPBS) to remove all traces of serum which contains trypsin inhibitor.
3. Add 5.0 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).4. Add 5.0 ml of complete growth medium and aspirate cells by gently pipetting.
5. Transfer all cell suspension to a 15ml centrifuge tube and spin at approximately 125 x g for 10 minutes. Discard supernatant.
6. Re-suspend the cell pellet in fresh growth medium. Add appropriate aliquots of the cell suspension to new culture vessels.
An inoculum of 3.0 X 104 to 5.0 X 104 viable cells/cm2 is recommended.
Incubate cultures at 37.0°C.
Storage temperature: liquid nitrogen vapor phase
Temperature: 37°C
