Sack: Thawing CHOcells

Sack and Yarov-Yarovoy Labs

Department of Physiology and Membrane Biology
University of California, Davis

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Thawing CHO Cells Jon Sack April 11, 2011 Ken Eum July 9, 2012 Basic Idea- Thaw fast in a happy place 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 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 Place tube of cells into the water bath for 1-2 min Thaw the cells quickly As soon as the cell are thawed, add 2 drops to 1 flask and the rest to the 2nd flask Place flasks into the 37° C incubator Change media 2 hours later with pen/strep No selection agents Change media 4-20 hours later, include pen/strep and selection agents Split cells before 80% confluent