I have a low quantity (~1 million) of human natural killer cells that I need to wash several times. These are non-adherent cells in suspension. I am currently spinning these cells down at 300xG for 10 minutes at 23C in a volume of ~4 mL in a 15 mL conical tube. However, I can't even really see a pellet. After aspirating the supernatant, resuspending, and doing a cell count, I am getting extremely low recovery (< 5%). I am likely losing a lot of cells when aspirating the supernatant, which I am currently doing with a vacuum. Any advice for better recovery would be appreciated. Thanks!
300g for 10mins should be enough to pellet your cells. I prefer to use eppendorfs or 5ml FACS tubes for smaller volumes as I can see the pellets more easily.
I also recommend aspirating with a pipette tip rather than vacuum until you find out where you are losing your cells.
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