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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
- 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
- Change media 4-20 hours later, include pen/strep and selection agents
- Split cells before 80% confluent
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