May 02, 2022
Version 1
Passaging of cells V.1
- 1AG Gerhardt
Protocol Citation: Guido Krähenbühl 2022. Passaging of cells. protocols.io https://dx.doi.org/10.17504/protocols.io.ewov1n3qkgr2/v1
License: This is an open access protocol distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
Protocol status: Working
We use this protocol and it’s working
Created: May 02, 2022
Last Modified: May 02, 2022
Protocol Integer ID: 61793
Abstract
General Lab procedure, Cell passage
Before start
Warm up culture media, PBS and Trypsin in a 37 °C Waterbath
Inspect cells for confluency, when 70-90 % confluency is reached cells are ready for passage
Discard media and wash once with PBS
Add Trypsin to fully cover the surface of the culture vessel
Incubate for 00:05:00 at 37 °C
5m
Lightly tap the culture vessel and check for cell detachement. If cells are still attached prolong incubation
Add 2/3 of culture media to 1/3 of trypsin and transfer cells into a Falcon tube
Centrifuge at 300 rcf, Room temperature, 00:05:00
5m
Discard supernatant and resubstitute with 1 mL culture media
Count cells and adjust media volume for required cell density
Plate cells