Oct 27, 2023

Public workspaceThawing of cells

This protocol is a draft, published without a DOI.
  • 1Charité
Bettina Ergün
Open access
Protocol Citation: Bettina Ergün 2023. Thawing of cells. protocols.io https://protocols.io/view/thawing-of-cells-c3ifykbn
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: October 17, 2023
Last Modified: October 27, 2023
Protocol Integer ID: 89383
Abstract
This method describes the process of thawing permanent cell lines
Materials
Growth medium
Falcon tube
T75 flask

Required
Cells
Required
Growth Medium

Allow cryotubes to thaw for Duration00:01:00 at RT at RT, then thaw for approx. Duration00:02:00 at 37 °C in a water bath while swirling until the last ice disappears

3m
Disinfect the outside of the tube with alcohol
Pipette the content into a Falcon tube containing 10 ml of pre-warmed growth medium
CAVE: In case of very sensitive cells, the cell suspension can be transferred from the cryotube directly into the culture bottle without centrifugation
Aspirate supernatant and resuspent pellet in growth medium (for T75: 4 ml )

Centrifigation300 x g, Room temperature, 00:04:00

4m
at the resuspented cell without bubbles to a culture flask prefilled with growth medium (for T75: 8 ml)

the size of the culture flask depends on the number of cells:
for 1-2E+6 cells => 1x T75