Review

Rev Diabet Stud, 2017, 14(1):22-38 DOI 10.1900/RDS.2017.14.22

Enzyme Development for Human Islet Isolation: Five Decades of Progress or Stagnation?

Daniel Brandhorst, Heide Brandhorst, Paul R.V. Johnson

Nuffield Department of Surgical Sciences, University of Oxford, United Kingdom
Address correspondence to: Daniel Brandhorst, Nuffield Department of Surgical Sciences, University of Oxford, John Radcliffe Hospital, Level 6, Headley Way, OX3 9DU, Oxford, United Kingdom, email: Daniel.Brandhorst@nds.ox.ac.uk

Manuscript submitted June 7, 2017; accepted June 9, 2017.

Keywords: diabetes, collagenase, islet isolation, neutral protease, thermolysin, tryptic-like activity, clostripain, extracellular matrix protein, basement membrane

Abstract

In comparison to procedures used for the separation of individual cell types from other organs, the process of human pancreatic islet isolation aims to digest the pancreatic exocrine matrix completely without dispersing the individual cells within the endocrine cell cluster. This objective is unique within the field of tissue separation, and outlines the challenge of islet isolation to balance two opposing priorities. Although significant progress has been made in the characterization and production of enzyme blends for islet isolation, there are still numerous areas which require improvement. The ultimate goal of enzyme production, namely the routine production of a consistent and standardized enzyme blend, has still not been realized. This seems to be mainly the result of a lack of detailed knowledge regarding the structure of the pancreatic extracellular matrix and the synergistic interplay between collagenase and different supplementary proteases during the degradation of the extracellular matrix. Furthermore, the activation of intrinsic proteolytic enzymes produced by the pancreatic acinar cells, also impacts on the chance of a successful outcome of human islet isolation. This overview discusses the challenges of pancreatic enzymatic digestion during human islet isolation, and outlines the developments in this field over the past 5 decades.

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