title: Endogenous pancreatic enzyme activity levels show no significant effect on human islet isolation yield.
abstract: Despite advances in human islet isolation, islet yield remains inconsistent and unreliable. In recent studies, it has been suggested that serine proteases, in particular trypsin, have been shown to have a damaging effect on islet yield. This study evaluated enzyme activity levels throughout 42 human islet isolation procedures. Trypsin, chymotrypsin, and elastase activity was determined spectrophotometrically using suitable chromophoric substrates. The results of the islet isolations were rated as successful (n = 19) or unsuccessful (n = 23) based on the islet yield and functionality. The enzyme activity profiles of the isolations were compared. No significant differences in donor-related variables were found in this study. However, in the successful isolations, a significantly greater amount (85.6 +/- 1.9%; p = 0.0017) of the pancreas was digested in a significantly shorter digestion time (19.7 +/- 0.6 min; p = 0.0054) compared with 74.8 +/- 2.5% of digested tissue in 22.6 +/- 0.7 min in the poor isolations. This study showed no significant effect of serine protease levels on the outcome of islet isolations, regardless of enzyme inhibitor supplementation. These data suggest that serine protease activity does not sufficiently affect islet yield. However, the data show that the most successful human islet isolations are achieved when the maximum amount of tissue is digested in the shortest amount of time. This suggests that further understanding of the isolation process should focus on the role of the collagenase digestion solution in the dissociation of the endocrine-exocrine tissue connection.
authors.affiliation: Surgical-Medical Research Institute, 1074 Dentistry/Pharmacy Centre, University of Alberta, Edmonton, Alberta, T6G 2N8, Canada.
authors.forename: Natisha L
authors.initials: NL
authors.lastname: Rose
authors.forename: Monica M
authors.initials: MM
authors.lastname: Palcic
authors.forename: A M James
authors.initials: AM
authors.lastname: Shapiro
authors.forename: Jonathan R T
authors.initials: JR
authors.lastname: Lakey
mesh_terms: D002469:Cell Separation; D002470:Cell Survival; D017364:Collagenases; D006801:Humans; D007515:Islets of Langerhans / Q000166:cytology / Q000201:enzymology*; D016381:Islets of Langerhans Transplantation; D012697:Serine Endopeptidases / Q000096:biosynthesis*
chemical_list: D012697:Serine Endopeptidases; D017364:Collagenases