I am growing cancer cells embedded in matrigel. I would like to split them into a new wells and continue growing them embedded in matrigel. Does anyone have a protocol for passing these cells? Should I use trypsin? Accutase? EDTA?
2hrs in Dispase (Becton Dickinson - now from LifeTechnologies) at 37°C will digest the Matrigel, how well your cells will tolerate that is hard to predict.
Thanks Nicholas - I'll try that. I'm using primary BCC cells. Nothing is really know about how they'll respond, so it's a crap shoot.
Use collagenase/disease (neutral protease). It will shorten the incubation time and can be done at 37oC.
Hi, we work with CRC organoids/spheroids and use the following procedure: 30-45 minutes in Cell Recover Solution (Corning) in a falcon tube on ice. When the organoids are visually on the bottom of the tube and/or after spinning form a nice compact pellet, wash with HBSS (or PBS).
Then if the cells can handle it ~5 minutes in trypsin (Sigma, diluted 1:1 with HBSS). Either inhibit the trypsin with medium+serum (wash out afterwards) or extensively wash out the trypsin before reseeding.
If cells don't like trypsin, try mechanical separation (syringing)...
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