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The QIAGEN Guide to Animal Cell Culture

IMPORTANT: Wear protective goggles and gloves when thawing vials that have been stored in liquid nitrogen. Vials may explode when removed from liquid nitrogen.

IMPORTANT: 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.

4. Wash the outside of the vial with 70% ethanol or another suitable disinfectant.

5. 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.

IMPORTANT: Thoroughly mix the cells in the cell culture vessel to ensure even distribution of the cells throughout the vessel.

6. Incubate cells overnight under their usual growth conditions.

7. The next day, replace the growth medium

Trypsinizing cells

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).

1. Aspirate the medium and discard.


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