Growing HEK293 EBNA1 cell line in suspension in Freestyle Media from Gibco. The cells seem to be having a tough time growing, increasing 20,000-40,000 a day. Even after starting a new and fresh culture the same results were observed. Papers don't seem to have anything on this so does anyone have any info on why this would be?
It could be the media, as it's not the suggested one, but a previous student was able to grow the cells fine in it many years ago (nothing in his notes about slow growth). Growing for transfection.
Also, after about a week or so of growth the cell count begins to decrease. Viability is still good. Not sure what's happening there either...
The first 2 things I would check for are Mycoplasma contamination or a bad batch of serum supplementing your media.
There's lots of kits available to test your cell lines for Mycoplasma in-house - it'd be worth giving one a try and consistently testing for it going forward if you don't already.
For serum, maybe try borrowing an aliquot from another lab or asking the supplier for a sample from a different lot. You could then try growing your cells in parallel and seeing if the different serum batches affect the growth rate.
Oops...my mistake. I didn't realize Freestyle media is serum free. You should still check for Mycoplasma though
the media as chemical formulated, is a sensitive one. check the expire date and whether you give the essential additives. (usually some should keep frozen and should be given just before use, etc.)
- Badges
- Science method