CRE BAG 2 producer cells are grown in Growth Media (GM, see Subheading 2.1.1., item 2) in a humidified 37°C incubator containing 5-7% CO2.
1. Remove a cryogenic vial from liquid nitrogen storage and rapidly thaw it in a 37°C water bath.
2. Transfer the contents of the vial to a 100-mm dish containing 10 mL of GM.
3. On the following day, aspirate the spent media and refeed the cells with 10 mL of fresh GM.
4. Feed the cultures with fresh GM every 2-3 d, subculture the cells 1:4 using trypsin/EDTA when cells reach approx 80% confluence (see Note 14).
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