Has anyone grown adherent cell lines (CHO-K and 293) in the Freestyle range of media's? Or another type of serum free media optimised for protein expression and purification?
Traditionally these Freestyle media by Gibco are designed for cells in suspension. I’m not entirely sure why adherent cells are not recommended, and Invitrogen can't provide much more information. So I was wondering if anyone has "bent the rules"?
And how do you find thier protein expression compares to media with serum?
My current problem is a contaminating protein in my purification which is suspect is from the serum
I've tried supplementing the media with ITS but the protein production suffers
As is always the case with these sorts of question, the answer depends on which cell types you are using and what you want to acheive with them. I have used a number of serum-free formulations for a number of different applications, and the exact requirements have varied greatly. When I tried to grow human hematopoietic stem cells in serum-free medium, I was unable to do so until I supplemented the medium with fibronectin. Some cell lines grow well in serum-free conditions, others not so. Many hybridoma lines grow just fine, but others suffer a drop in antibody yield.
We currently grow Hek 293 cells in freestyle media. We directly adapted the cells by adding them into freestyle media at 1 x 106/ml, you may see a loss in viability but they do recover, just change media every 2-3 days and keep them at high density (1 x106/ml). Even in freestyle they are very sticky if left alone without agitation and will settle down over time.
regards
Ian
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