I sorted for Jurkat cells which express CD25 surface marker and was able to get around 300,000 to 400,000 cells. I resuspended them in 5 ml of RPMI media with 10% FBS and 1% pen-strep. After two days, the cells look shrunk and it's not the usual spherical/round shaped-cells. The thing is I didn't check how the cells look like after sorting since I was running out of time for another experiment. Do you guys know if this usually happens after sorting and do you think the cells would be able to recuperate? Thanks!
Is the collection tube coated with FBS, before the cell are collected into the tube?. if not the cells will just adhere to the tube wall due to electrostatic interaction and cell lost will be high.
Hi Nagarjun, Thanks for your thoughts.
I used 15 ml falcon tube with 2 ml of RPMI media (with 10%FBS) as a collection tube. I still got some ample amount of cells when I resuspended it in the media but I'm just worried because after two days, they seemed to be shrunk and not in their normal morphology.
Thanks,
Carla
Update:
A week after sorting, the cells appear to be recuperating. My mentor said it usually takes a while for them to grow after sorting. We'll see how it goes.
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