Collagenase Type VIII From Sigma Aldrich for Tissue Digestion

Results Summary

We have been using Collagenase type VIII from C Histolyticum for my gut prep and immune cell isolation. We use this enzyme at a concentration of 1 mg/ml and digest a single colon in 25-30 ml of digestion media. Before putting the colonic tissue in the digestion media, we thoroughly wash the tissue in cold 1x PBS to remove the trace amount of EDTA, if used in the previous steps. The tissue is then finely chopped using the a sharp scissors and then 25-30 ml digestion media was added and kept at 37 degree C for 25-30 mins. The enzyme is very good and after 25mins incubation the entire tissue is generally digested. If at all small chunks left just mash it through a 70 micron strainer. We do not recommend incubating the tissue over half an hour as it increase the cell death.