I am trying to freeze attached cells and cannot use trypsin. Is there any other good method?
We have good results with accutase. It is gentler and do not cleaves surface markers.
http://www.accutase.com/accutase.html
You can try PBS with EDTA. If that is not enough you can use socalled cell stripping solutions (sold for instance by SIGMA or Corning).
Good luck ;)
Hi Tommye,
Do you need to freeze life cells to reculture them later or your trying to extract something from them?
-ED
You can also use citrate saline (135 mM potassium chloride, 15 mM sodium citrate), with or without 10 mM EDTA, depending on how adherent your cells are. I use this to remove cells so that I can collect the deposited extracellular matrix, but I have to admit that I've never re-plated the cells to check their viability.
In addition above, cell scrapers are commercially available to detach the adherent cells. NLS
Hi there!
If you want your cells as untouched/damaged as possible, I would go for Accutase. They keep the membrane integrity better than Trypsin or PBS+EDTA (at least in my experience). Cell scrapers might cause damage, but it also depends on how resistant your cells are ;)
Best!
I use this product, works good and simple
https://www.thermofisher.com/order/catalog/product/13151014
We are developing a cell culture coating that is thermoresponsive; cells attach at incubator temperatures, grow and then detach when vessels are allowed to cool.
https://www.isurtec.com/products/isurtherm/
More labor-intensive methods can involve spraying the cell layer with a pasteur pipet, or even scraping them off, but these are less gentle as well.
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