Dear Wormy people
Does anyone has good experience with embryonic cell dissociation? I am trying to dissociate the embryonic cells into single cells. But lots of them are still in clumps. I want to increase the yield, does anyone has tricks to maximize
the yield of dissociation.
Any help or suggestion would be greatly appreciated.
Thanking
UGANGIS
My grad lab used to use syringe filters to break up the big clumps. As stated in the embryonic cell culture section of Wormbook’s Methods in Cell Biology section, specifically protocol 32:
“Preload 3cc syringe with 0.5 ml media and draw up cell suspension into barrel. Gently push sample through a 5 μm syringe filter into a 1.5 ml microfuge tube. To recover more embryonic cells, reload the syringe with ~1.5 ml media and push it through the filter again into a second microfuge tube. The filtration step is crucially important for removing cell clumps and undissociated embryos.”
You can email David Miller to see if they have any updates, since this protocol was written in 2006.
Best of luck!
Steve V