Wear protective goggles and gloves when thawing vials
that have been stored in liquid nitrogen. Vials may explode when removed
from liquid nitrogen.
Proceed to step 4 as soon as the cells have thawed. Do
not allow the cells to warm up before transferring them into growth medium.
Wash the outside of the vial with 70% ethanol or another suitable
Slowly pipet the thawed cell suspension into the cell culture
vessel containing prewarmed growth medium. Swirl the vessel gently to mix
the cells with the medium.
Tip - Immediate removal of DMSO may sometimes be necessary, especially for
suspension cells, primary cells, and sensitive cell types. For such cell
types, pipet the thawed cell suspension into a sterile centrifuge tube containing
prewarmed medium, centrifuge at 200 x g for 2 min, aspirate the supernatant,
resuspend the cells in fresh growth medium, and then transfer to an appropriate
cell culture vessel.
Thoroughly mix the cells in the cell culture vessel to
ensure even distribution of the cells throughout the vessel.
Incubate cells overnight under their usual growth conditions.
The next day, replace the growth medium
Trypsinization is a technique that uses the proteolytic enzyme trypsin to
detach adherent cells from the surface of a cell culture vessel. This procedure
is performed whenever the cells need to be harvested (e.g., for passaging,
counting, or for nucleic acid isolation).
Aspirate the medium and discard.
Page: All 1 2 3 4 5 6 Related biology technology :1
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