I've coated T75 flasks in 0.1mg/mL poly-D-lysine but when I went to use accutase to detach them they wouldn't budge. Tried putting them in the incubator for a couple of hours and nothing, wondered if I should try trypsin or if it would be too harsh for these type of cells or if my concentrations weren't right?
Malcolm Nobre
Hello Roisin
There is no harm in using trypsin. You can use 0.25% Trypsin-0.53mM EDTA solution for a maximum of 5 minutes at 37 deg. Trypsin being a proteolytic enzyme will cleave proteins that help in cell attachment and EDTA being a metal chelator will chelate Ca2+ and Mg2+ in the extracellular matrix especially Cadherins which help in cell-cell adhesion. EDTA will help trypsin to act more efficiently.
Good Luck.
0 votes
0 thanks
- Badges
- Science topic
- Similar topics
- Comment
More Roisin Moloney's questions See All
Similar questions and discussions