I am culturing iPSCs on matrigel coated plates using mTeSR media. When I manually lift off iPSC colonies for neurosphere formation with the intent of further differentiation into neurons I get a polymer that forms in the cell suspension. Usually I notice it after a day and looks like big globs of polymer that engulf all my newly formed spheres. Any suggestions on what is causing this and what can be done to prevent it?
Hi! It could be matrigel solution. How you prepape your matrigel?did you see any precipitate or any gelification from your matrigel when you precoat the plates?you will see them in your plates as small polymer or scaffold of matrigel. It will affect your ipsc differentiation too. How you lift cells up?usually we use dispase or Releserd for the ipsc. I hope it could help. Miguel
Hi,
I also think it is a matrigel issue, how do you do the coating? Matrigel gets gellified very easily unless kept on ice, including the tips used for pipetting it. Also, for lifting up the colonies, I would recommend using ReLeSR, which nicely loosens the colonies.
If the problems still remains, you might also try vitronectin instead of matrigel, which I find much easier to work with and have not observed any difference in culture quality compared to matrigel.
Good luck!
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