Sack: Thawing CHOcells

Thawing CHO Cells Jon Sack April 11, 2011
Ken Eum July 9, 2012

Basic Idea- Thaw fast in a happy place

1. Add 5ml of cell media (F12 + 10% FBS) without antibiotics or selection agent to two 25cm2 flasks and place into 37° C incubator for 1 hour
2. Remove cells from liquid nitrogen freezer
   • make sure to transport on dry ice and note removal in the liquid nitrogen log
   • KEEP THE CELLS FROZEN ON DRY ICE
3. Place tube of cells into the water bath for 1-2 min
   
   • Thaw the cells quickly
4. As soon as the cell are thawed, add 2 drops to 1 flask and the rest to the 2nd flask
5. Place flasks into the 37° C incubator
6. Change media 2 hours later with pen/strep
   • No selection agents
7. Change media 4-20 hours later, include pen/strep and selection agents
8. Split cells before 80% confluent

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