Hi all, after culturing hepatocytes in a monolayer on collagen in 24 well plates for 5 days, I attempted to trypsinize (0.25% trypsin) them to perform a cell count. However, none of my cells lifted off the plate. I let them sit in trypsin for two hours out of curiosity and they wouldn't budge.
Do I need to use collagenase as well? Do I need to use both collegenase and trypsin? I do necessarily need a single cell suspension, but I do need to perform an accurate cell count.
Hello!
Please you look at
Article Recent advances in the isolation of liver cells
The perfusate is changed to a solution containing a combination of proteolytic enzymes (e.g., 0.02% collagenase with 40 to 80 kg/ml DNase I and incubated for 20 min at 37" C.
You will need collagenase. Trypsin will not be needed. Article Subculture of proliferating adult rat hepatocytes in medium ...
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