Passaging of cells
Guido Krähenbühl
Published: 2022-05-02 DOI: 10.17504/protocols.io.ewov1n3qkgr2/v1
Abstract
General Lab procedure, Cell passage
Before start
Warm up culture media, PBS and Trypsin in a 37°C
Steps
1.
Inspect cells for confluency, when 70-90 % confluency is reached cells are ready for passage
2.
Discard media and wash once with PBS
3.
Add Trypsin to fully cover the surface of the culture vessel
4.
Incubate for 0h 5m 0s at 37°C
4.1.
Lightly tap the culture vessel and check for cell detachement. If cells are still attached prolong incubation
5.
Add 2/3 of culture media to 1/3 of trypsin and transfer cells into a Falcon tube
6.
Centrifuge at 300rcf
7.
Discard supernatant and resubstitute with 1mL
8.
Count cells and adjust media volume for required cell density
9.
Plate cells
