Two days ago I thawed a freezer stock of HEK293T cells into a fresh batch of sterile media. I can clearly see some normal-looking cells adhered to the plate, but there are also large numbers of floating cells. These seem to be growing, as you can see them mid-division (bottom left of photo), but they are also seen in long chains that are reminiscent of yeast. The size, in comparison to the adherent HEKs, makes me think they are larger than typical yeast contaminants, and the observed division doesn't look like the budding I'd expect from yeast. I would expect a large number of dead cells to be present in suspension in a culture recovering from a freezer stock, but I wouldn't expect to see the floating cells dividing. I'm about to change out the media as well as put some of my fresh media in a plate to see if anything grows. Based on the picture and my observations so far, what do you think? I'll update with my findings re: fresh media.
HI Eric,
In my experience, HEK293T cells are capable of growing in suspension if for some reason they fail to attach.
I am growing my HEK293T cells in Freestyle-293-expression-medium and they grow in suspension in this medium. This medium is serum free, so my guess is that low serum may cause cells to remain in suspension.
On the other hand the culture surface may also cause the cells to remain in suspension, for example the use of cell-culture non treated plats.
Another possibility could be that cells are still recovering from the freeze shock and need more time to attach.
Good luck,
Sudeep
Sudeep Kumar, do your HEK293T cells growing in suspension look like those in my picture?
Yes, they look fine to me.
Once they grow more, they will make bunches though, if they are still in suspension.
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