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BACKGROUND: The isolation of islets from the human pancreas critically depends on an efficient enzyme blend. Previous studies have solely focused on the presence of collagenase and neutral protease/thermolysin. Despite improved characterization of these components, the lot-related variability in efficacy still persists suggesting that additional so far disregarded enzymes are required for efficient islet cleavage. METHODS: Varying activities of a tryptic-like enzyme were identified within collagenase NB1 lots, which were selected according to a matched ratio between tryptic-like and collagenase activity (TLA-ratio). Rat and human pancreata were processed with current standard procedures. RESULTS: Increasing the TLA-ratio from 1.3% to 10% reduced pancreas dissociation time in rats by 50% without affecting islet yield, viability, or posttransplant function in diabetic nude mice. Enhancing the TLA-ratio from 1.3% to 12.6% for human pancreas processing resulted in a significant reduction of recirculation time and increased incrementally human islet yield without affecting purity, in vitro function or recovery after culture. Optimized pancreas digestion correlated with a higher percentage of islet preparations fulfilling quality criteria for clinical transplantation. CONCLUSIONS: We conclude that TLA is an effective component that should be included in moderate amounts in enzyme blends for human islet isolation to optimize the efficiency and minimize the lot-related variability.

Original publication

DOI

10.1097/TP.0b013e31819499f0

Type

Journal article

Journal

Transplantation

Publication Date

15/02/2009

Volume

87

Pages

370 - 375

Keywords

Animals, Cell Separation, Cell Survival, Collagenases, Diabetes Mellitus, Experimental, Female, Graft Survival, Humans, Insulin, Insulin-Secreting Cells, Islets of Langerhans, Islets of Langerhans Transplantation, Male, Mice, Mice, Nude, Middle Aged, Rats, Rats, Inbred Lew, Thermolysin, Tissue Donors, Transplantation, Heterologous, Trypsin