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Chapter: Basic Concept of Biotechnology - Animal Biotechnology

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Protocol for Thawing Frozen Cells

The following protocol describes a general procedure for thawing cryopreserved cells. For detailed protocols, always refer to the cell-specific product insert.

Thawing Frozen Cells

 

Protocol for Thawing Frozen Cells

 

The following protocol describes a general procedure for thawing cryopreserved cells. For detailed protocols, always refer to the cell-specific product insert.

 

1.        Remove the cryovial containing the frozen cells from liquid nitrogen storage and immediately place it into a 37oC water bath.

 

2.        Quickly thaw the cells (< 1 minute) by gently swirling the vial in the 37oC water bath until there is just a small bit of ice left in the vial.

3.        Transfer the vial it into a laminar flow hood. Before opening, wipe the outside of the vial with 70% ethanol.

 

4.        Transfer the thawed cells drop wise into the centrifuge tube containing the desired amount of pre-warmed complete growth medium appropriate for your cell line.

 

5.        Centrifuge the cell suspension at approximately 200~ g for 5–10 minutes. The actual centrifugation speed and duration varies depending on the cell type.

 

6.        After the centrifugation, check the clarity of supernatant and visibility of a complete pellet. Aseptically decant the supernatant without disturbing the cell pellet.

 

7.        Gently re-suspend the cells in complete growth medium, and transfer them into the appropriate culture vessel and into the recommended culture environment.

 

Note: The appropriate flask size depends on the number of cells frozenin the cryovial, and the culture environment varies based on the cell and media type.


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