FreeStyle™ 293 Expression Medium

With FreeStyle 293 Expression Medium, we would recommend harvesting the cells a little earlier to reduce any nonspecific binding by host cell proteins and then filtering the culture supernatant through a 0.2 or 0.1 µM membrane before loading onto the column. On the other hand, FreeStyle CHO Expression Medium contains EDTA that will strip the nickel column. The medium can be subjected to dialysis or buffer exchange prior to loading on the column, or the EDTA can be chelated by adding 1-3 mg/L NiSO4 or NiCl to the supernatant. Iron or cobalt salts can also be used.

You should be able to use the ExpiFectamine293 Reagent for transfection of Freestyle 293-F cells grown in FreeStyle 293 Expression Medium; however, the enhancers will provide only a little boost in expression as they are designed to work with higher density cultures that FreeStyle 293 Expression Medium cannot support.

Find additional tips, troubleshooting help, and resources within our Transfection Support Center.

Our recommendation of 8% CO2 for the optimal growth of Freestyle 293 cells in Freestyle 293 Expression Medium is based on the bicarbonate content in the media and growth conditions of the cells observed by R&D. That said, you may use lower concentrations of CO2 if necessary (5% is common) to grow these cells. You may observe slightly slower growth kinetics or lower cell concentrations, but functionally the entire system will still work.

Yes, the FreeStyle 293 Expression Medium contains Pluronic F-68 and lipids that would interfere with ammonium sulfate precipitation. Dialysis or tangential flow filtration can be used to remove lipids and Pluronic F-68 from the medium. Then you can concentrate the medium with ammonium sulfate.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.

FreeStyle F17 Expression Medium was designed from a different base formulation than FreeStyle 293 Expression Medium. FreeStyle F17 medium shows equal or better performance than the FreeStye 293 medium for certain cell lines and proteins. It is capable of supporting the growth of 293 and CHO cells and offers a wider variety and selection of transient cell culture media options.

Find additional tips, troubleshooting help, and resources within our Cell Culture Support Center.