Indexed on: 19 Oct '17Published on: 19 Oct '17Published in: American Journal of Transplantation
High number of human islets can be isolated using modern purified tissue dissociation enzymes, however it requires using >20 Wunsch Unit (WU)/gram of pancreas for digestion. Attempts to reduce this dose have resulted in pancreas under-digestion and poor islet recovery but improved islet function. In this study, we achieved high number of functional islets using low dose of recombinant collagenase enzyme mixture (RCEM). The collagenase dose used in these isolations is about 42% of the natural collagenase enzyme mixture (NCEM) dose commonly used to digest a 100g pancreas. Low dose RCEM was efficient in digesting entire pancreases to obtain higher yield (5,535±830 IEQ/gram and 2,582±925 IEQ/gram, P<0.05) and lesser undigested tissue (16.7±5% and 37.8±3%, P<0.05) when compared to low dose NCEM (12WU/gram). Additionally, low dose RCEM islets retained better morphology (confirmed by scanning electron microscopy), and higher in vitro basal insulin release (2391 ± 1342 μU/ml and 1778 ± 978 μU/ml; P<0.05) when compared to standard NCEM dose. Nude mouse bioassay demonstrated better islet function for low dose RCEM (AUC 24,968) over low (AUC-38,225) or standard NCEM dose (AUC-38,685), p<0.05. This is the first report indicating that islet function can be improved by using low dose rC1rC2 (RCEM). This article is protected by copyright. All rights reserved.