BACKGROUND: Efficient islet isolation represents a necessary requirement for successful islet transplantation as a treatment for type 1 diabetes. The choice of collagenase for pancreas digestion is critical for the isolation outcome, and Liberase is the most widely used enzyme, although large intra-batched variability in activity and efficiency has been observed. METHODS: The aim of this study was to characterize Liberase components and their relative role in pancreas digestion. Liberase batches were characterized by microelectrophoresis. RESULTS: By means of microelectrophoresis, we identified three main proteins each with different prevalences between batches. Two proteins were found to correspond to class I (CI) and one to class II (CII) collagenase. In a series of 163 islet isolations, we observed that the CII correlated with islet yield (P<0.001) and digestion time (P<0.001); additionally, CI directly correlated with purity (P=0.028). Finally, when CII and one of the CI isoforms were >50 percentile, 15 of 36 preparations were transplanted, with 27 of 127 transplanted in the other cases (P=0.013). CONCLUSION: These results represent an important step toward the characterization of enzymes, with the final aim of identifying key components for a standardized product.
BACKGROUND: Efficient islet isolation represents a necessary requirement for successful islet transplantation as a treatment for type 1 diabetes. The choice of collagenase for pancreas digestion is critical for the isolation outcome, and Liberase is the most widely used enzyme, although large intra-batched variability in activity and efficiency has been observed. METHODS: The aim of this study was to characterize Liberase components and their relative role in pancreas digestion. Liberase batches were characterized by microelectrophoresis. RESULTS: By means of microelectrophoresis, we identified three main proteins each with different prevalences between batches. Two proteins were found to correspond to class I (CI) and one to class II (CII) collagenase. In a series of 163 islet isolations, we observed that the CII correlated with islet yield (P<0.001) and digestion time (P<0.001); additionally, CI directly correlated with purity (P=0.028). Finally, when CII and one of the CI isoforms were >50 percentile, 15 of 36 preparations were transplanted, with 27 of 127 transplanted in the other cases (P=0.013). CONCLUSION: These results represent an important step toward the characterization of enzymes, with the final aim of identifying key components for a standardized product.
Authors: A N Balamurugan; Gopalakrishnan Loganathan; Melena D Bellin; Joshua J Wilhelm; James Harmon; Takayuki Anazawa; Sajjad M Soltani; David M Radosevich; Takeshi Yuasa; Mukesh Tiwari; Klearchos K Papas; Robert McCarthy; David E R Sutherland; Bernhard J Hering Journal: Transplantation Date: 2012-04-15 Impact factor: 4.939
Authors: Orison O Woolcott; Richard N Bergman; Joyce M Richey; Erlinda L Kirkman; L Nicole Harrison; Viorica Ionut; Maya Lottati; Dan Zheng; Isabel R Hsu; Darko Stefanovski; Morvarid Kabir; Stella P Kim; Karyn J Catalano; Jenny D Chiu; Robert H Chow Journal: Pancreas Date: 2012-01 Impact factor: 3.327
Authors: Magnus Ståhle; Aksel Foss; Bengt Gustafsson; Marko Lempinen; Torbjörn Lundgren; Ehab Rafael; Gunnar Tufveson; Olle Korsgren; Andrew Friberg Journal: Transplant Direct Date: 2015-06-24