i am trying to seed cells on coverslip coated with gels (acrylamide based hydrogels)
My main obstacle is my cells aren't settling down as expected so they can grow instead they keep floating, or migrating to the edges of the gel and there's almost no cells in the center of my gel.
Any ideas on how to solve this dilemma?
Your cells do not like it as matrix. Do you prepare it yourself? As i see after a quick check in the literature (i never heard this before, now i learned something) the polymer is decorated with ECM proteins for cells.
Yes I do prepare the gel myself and also incubate the gel overnight @ 4 degrees with collagen prior to cell seeding.
I do not check the details for your protocol, but but collagen has to be prewarmed before using to pretreat... and in your case (because of the 3D-matrix), the diffusion even slower the process. leave the gel and collagen at longer time at RT at least, or maybe copoymerize them... Good luck!
In addition to the ECM protein recommendations, you might consider plasma treating (air or oxygen) the gel to increase its hydrophilicity. 1-2 minutes should be fine.
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