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Comparison of Neutral Proteases and Collagenase Class I as Essential Enzymes for Human Islet Isolation
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology. Univ Oxford, Nuffield Dept Surg Sci, Oxford, England.;Oxford Ctr Diabet Endocrinol & Metab, Oxford, England..
Serva Electrophoresis GmbH, Uetersen, Germany..
Univ Oxford, Nuffield Dept Surg Sci, Oxford, England.;Oxford Ctr Diabet Endocrinol & Metab, Oxford, England.;Oxford NIHR Biomed Res Ctr, Oxford, England..
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology, Clinical Immunology.ORCID iD: 0000-0002-8524-9547
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2016 (English)In: TRANSPLANTATION DIRECT, ISSN 2373-8731, Vol. 2, no 1, article id e47Article in journal (Refereed) Published
Abstract [en]

Background. Efficient islet isolation requires synergistic interaction between collagenase class I (CI) and class II (CII). The CI degradation alters the ratio between CI and CII and is responsible for batch-to-batch variations. This study compares the role of neutral protease (NP) plus clostripain (CP) with CI as essential enzymes for human islet isolation.

Methods. Human islets were isolated using 4 different enzyme mixtures composed of CII plus either intact (CI-115) or degraded CI (CI-100). Blends were administered either with or without NP/CP. Purified islets were cultured for 3 to 4 days before islet quality assessment.

Results. Whereas using intact CI-115 without NP/CP did not significantly reduce islet yield (3429 +/- 631 vs 3087 +/- 970 islet equivalent/g, nonsignificant), administration of degraded CI-100 without NP/CP decreased islet yield from 3501 +/- 580 to 1312 +/- 244 islet equivalent/g (P < 0.01), doubled the amount of undigested tissue from 11.8 +/- 1.6 to 24.4 +/- 1.2% (P < 0.01) and triplicated the percentage of trapped islets from 7.7 +/- 2.8 to 22.5 +/- 3.6% (P < 0.05). Islet yield did not vary between supplemented CI-115 and CI-100, but was increased using CI-115 when NP/CP was omitted (P < 0.05). A trend toward higher viability and increased secretory insulin response was noted in both CI-100 and CI-115 when NP/CP was not added.

Conclusions. This study suggests that NP/CP can compensate reduced CI activity. Future attempts to optimize enzyme blends should consider the possibility to increase the proportion of collagenase CI to reduce the need for potentially harmful NPs.

Place, publisher, year, edition, pages
2016. Vol. 2, no 1, article id e47
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Medical and Health Sciences
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URN: urn:nbn:se:uu:diva-313439DOI: 10.1097/TXD.0000000000000552ISI: 000390125900002PubMedID: 27500241OAI: oai:DiVA.org:uu-313439DiVA, id: diva2:1069810
Available from: 2017-01-30 Created: 2017-01-19 Last updated: 2017-01-30Bibliographically approved

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