Hi,
I am currently working on my master thesis and I want to do a transwell migration assay with bone marrow derived macrophages in January. I have aliquots of the macrophages in liquid nitrogen storage and when I use them for different experiments, I usually let them recover for about 2 days with daily medium changes. The harvest (I scrape them off the dish) and the freezing-thawing process puts a lot of stress on them, so I like to give them this time of recovery and get rid of dead cells.
When I searched for protocols for the migration assay, I only find that the cells are grown in another dish prior to migration and then detached by Trypsin and seeded into the transwells for the migration. I would like to go around this detachment-step, as it would be additional stress for the macrophages. So could I also seed the macrophages directly into the transwells (5 µm pore-size) after thawing them and let them recover in there (including medium changes) or would they already start migrating even without a stimulus? Or is there any other way how I could go around the stress of the additional detaching?
Thank you for your help!
Hi Jana,
I have the same doubt and just wanted to check with you whether you find the answer for this question?
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