Hello!
I have mouse liver spheroids (generated in vitro in 96-well plates for 10 days, they measure around 200 um) and I want to dissociate them into live single cells for FACS experiment.
So far I have tried:
- Accutase, 37C with shaking at 500 rpm for 10 min. If I incubate for less time the spheroids don't desintegrate, but when they finally fall apart and I stain them with Trypan they are all dead.
- Liberase (Roche, low thermolysin concentration) at 25 ug/ml - this is the concentration I use to isolate the hepatocytes from the liver, so although it may not be the perfect concentration, I thought I would be a good place to start. Again, incubation at 37C, shaking for 10 min, they take a while to fall apart, but when they do and I stain with Trypan, they are all dead.
Has anybody had any success with this, could you please help me out??
Thanks in advance!!
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