Recommended Cell Culture Conditions:

Cell Recovery: Thaw cells in a 37 °C water bath with shaking until the mixture has dissolved. Transfer to a centrifuge tube and add culture medium (see Recommended Cell Culture Conditions above) at a volume 3-5 times the volume of the cells. Centrifuge at 1000 RPM for 5 minutes and discard the supernatant. Transfer to a T25 flask for culture.

Suggested Cell Passage Procedure: Cells should be 85-95% confluent before cell passage is carried out.

1. Discard the medium and wash with PBS 1-2 times.
2. Add 1 ml of Trypsin at 37 °C, then observe the cells under a microscope.
3. When the cells appear retracted and rounded, gently tap the culture flask to detatch the cells. Stop the trypsinization by adding 2 ml of culture medium containing 10% serum.
4. Add fresh medium to resuspend the cells. The recommended ratio of primary cells is 1/2. Pipette to obtain a single cell suspension.