Hello everyone,
I am having trouble with getting rid of old matrigel fragments when I passage my organoids from an old dome to a new one. I see that some organoids remain stuck in old matrigel fragments that do not get incorporated into the new dome. Unfortunately, the corning matrigel cell recovery solution is currently unavailable. I have read that some people use Dispase solution. Has anyone tried this? If yes, which dispase? Or does anyone have any other tips?
Thanks a lot in advance!
Hello Claudia Owczarek
For recovering organoids from Matrigel matrix, the culture may be treated with Corning Cell Recovery Solution. You should not use Dispase as it will make single cell suspension, unless this is what you want.
Best.
Hi Malcolm Nobre ,thank you for your answer. As I mentioned, the Corning Cell Recovery Solution is currently unavailable because of an ongoing shortage. Do you know of any alternatives that can be used?
Thanks again!
It depends on what your downstream application is?
Corning cell recovery solution works very well, but is expensive and takes time.
An alternative would be to gather organoids and matrigel in ice cold media or dpbs, which will liquify the matrigel again. If gather lots of domes place on ice for longer (5mins). Spin down and aspirate solution, matrigel will sit on top of the organoids.
See Article Establishment and Culture of Human Intestinal Organoids Deri...
for an excellent detailed description.
Hi Malcolm Nobre How to use the corning cell recovery solution to the organoids? Can you share the protocol?
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