Cell freezing protocol for cells growing in log phase

Cell freezing protocol:

Exponentially growing cells (log phase) can be stored frozen for many years using cryoprotective agent such as DMSO and glycerol.

1. Trypsinize subconfluent cells with 1x Trypsin medium for few minutes, watch cells round off and detach from bottom. Stop reaction by adding 1 ml fetal calf serum or calf serum.

2. Centrifuge at 1000 rpm for 5 min and resuspend cells in appropriate volume of freezing media (10% DMSO, 90% FCS) at 2-4 x 10^6 cells/ml.

3. Transfer cells resuspended in freezing medium to a sterile cryogenic vial, label vial clearly (date, cell line name, cell concentration etc).

4. Put cryogenic vial in freezing rack and store in -80c for 24 h before transfering to liquid nitrogen tank for long-term storage.

Last update 25-Jul-2006, Rating Very Good of 2 votes.