I thaw the EA.hy cells from liquid nitrogen but they do not attach even 24hrs post seeding, those few cells that attached are growing very slowly. would you please help me to find a solution.
I am not really familiar with these cells. But anyways, here are some general tips that might help. Freezing in more FBS gives better yield after thawing, a friend of mine freezes his cells in just FBS with no medium Personally I don't go this far, I use 20% FBS. Freeze slow, thaw fast. When thawing try to thaw as fast as you can. When I thaw my cells, I warm them up in my hand, add worm medium, place the thawed cells in 10 ml medium. Mix up and down, then spin down cells. I remove medium and add new medium, mix gently up and down (I need not to say, never vortex your cells, this is ABC cell culture, yet in some labs it is done). count your cells, and seed them in a 25cm flask if the count is low. If the cells having hard time attaching you might consider using collagen. I hope this helps. This information is very basic, but just in case.