Take the cryovial from nitrogen, clean it with disinfectant, put it in the waterbath set to 37°C.

In the meanwhile prepare a flask tagging it with cell line name, passage, date and operator. Fill it with medium 10ml.

If coating is required, check in advance which kind of is needed. Perform the coating accordingly to the related procedure and then perform step 2.

Once vial is almost completely thawed, remove it from the waterbath. Prepare a falcon (cell line name, passage) with enough medium/Stop Solution to wash the cells.

Take the content of the cryovial and add it to the falcon. Wash the cryovial with 1 ml of medium/stopping solution.

Add it to the falcon.

Centrifuge cells at 1200 RPM for 5 minutes. Remove pellet, resuspend cells accordingly to the number of flask to the seeded and cells number. thorougly pipette to prevent aggregation and seed cells in the flask.