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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