The Basic steps of the cell-reviving process

The Basic steps of the cell-reviving process 

1)  Cryopreserved Cells [with DMSO (Dimethyl sulfoxide)] were collected from the -80°C.

2) Then the cells were thawed rapidly in the water bath at 37°C for 30-45 sec to protect the cells from DMSO.

3) The cells were transferred into a 15 ml falcon which already contains 6 ml of media [DMEM (Dulbecco’s Modified Eagle Medium) + 10% FBS (Foetal Bovine Serum) + 1X penicillin-streptomycin solution].

4) The centrifugation was performed at 1100 rpm for 5 minutes.

5) The Supernatant was discarded in the beaker containing a small amount of bleach and water. The pellet was collected.

6) The pellet was washed with media to remove the trace of DMSO.

7) The cells are resuspended in 1 ml of media. 

 8) The cells were transferred to the T-25 flask which already contains 4 ml of media. 

9) Finally, the cells were observed under the microscope and kept in the incubator at 37°C, with 5% CO₂ and humid conditions.