hello

is there a reason for HUVEC not adhering to the denuded amniotic membrane?

I have read several protocols that are quite similar except for the chemical used to remove the epithelial cells e.g. Na4OH, EDTA, trypsin, thermolysin and dH2O.

I am using trypsin/EDTA 0.5g/0.2g for 20 minutes at 37C then scraping the cells to ensure the epithelial cells will not interfere with HUVEC attachment. however, my HUVEC won't stick

could someone help explain why?

or maybe share a protocol which has been personally tried

I appreciate your support