im currently working on podocyte cell lines. protocol recommends use of accutase to detach adherent cells. can this be replaced by trypsin?
I am not experienced at working with Podocytes personally. However, over the past decade+, many people have found that Accutase works very well for certain cell types that may not be detached well by Trypsin. Accutase is a cocktail of cold-active proteases extracted (if I remember correctly) from the gut of crustaceans (the by-product of a canned crab-meat factory in San Diego or somewhere similar) and most companies that sell Accutase license or source the product from this one origin. This means that they can be more effective at detaching cells that have different extracellular matrix-components than those digested by Trypsin, and can also be more gentle as it is active at lower temperatures. It is also relatively cheap, so there is no good reason to not buy some (most of the usual companies cell it - most people here use the Invitrogen stuff from memory).
Trypsin *may* work ; however, you may find that it is either too harsh for your cell type, or simply not effective owing to a different matrix being secreted.
Best of luck.
I am not experienced at working with Podocytes personally. However, over the past decade+, many people have found that Accutase works very well for certain cell types that may not be detached well by Trypsin. Accutase is a cocktail of cold-active proteases extracted (if I remember correctly) from the gut of crustaceans (the by-product of a canned crab-meat factory in San Diego or somewhere similar) and most companies that sell Accutase license or source the product from this one origin. This means that they can be more effective at detaching cells that have different extracellular matrix-components than those digested by Trypsin, and can also be more gentle as it is active at lower temperatures. It is also relatively cheap, so there is no good reason to not buy some (most of the usual companies cell it - most people here use the Invitrogen stuff from memory).
Trypsin *may* work ; however, you may find that it is either too harsh for your cell type, or simply not effective owing to a different matrix being secreted.
Best of luck.
Robin gave an excellent answer, so I will only add that if you do in fact find trypsin to be too harsh, then you simply can dilute it with HBSS (without Ca or Mg) and/or shorten its exposure time.
- Badges
- Science topic