Collagenase and neutral protease1 are essential tissue dissociation enzymes (TDEs) required to release islets2 and other cells3-6 from tissue. The focus of many reports has been on the characteristics and performance of collagenase in islet isolation7,8 but a recent report by Brandhorst et al. showed that the presence of trypsin like activity (TLA) in selected lots of Serva purified collagenase when added to Clostridium histolyticum neutral protease correlated with higher human islet yields9. This post emphasizes the assays used to assess neutral protease activity present in tissue dissociation enzymes. It is the first of an ongoing series reviewing biochemical characteristics of these enzymes used in cell isolation procedures.
Archive for October, 2009
Tissue Dissociation Enzyme Neutral Protease Assessment
Wednesday, October 7th, 2009What is the appropriate assay to assess collagenase prior to use in cell isolation procedures?
Wednesday, October 7th, 2009The quality of collagenase containing tissue dissociation enzymes (TDEs) has a direct impact on the success of the cell isolation procedure (e.g., hepatocytes¹, islets²). Subsequent studies showed that both collagenase and a neutral protease were required to effectively dissociate cells from tissue. In the past, biochemical assays used to characterize the product served as a guide for purchase but the ultimate test was to assess the effectiveness of TDE product in a cell isolation procedure. VitaCyte’s characterization of purified class I (C1) and class II (C2) collagenase enzymes from Clostridium histolyticum by different biochemical assay procedures have provided useful comparative data for the most appropriate assays to characterize collagenase activity in any collagenase containing TDE products. The brief overview below provides an analysis of our current understanding of the pros and cons of different biochemical assays for collagenase.
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