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Updated: Mar 16, 2026
Mouse Islet of Langerhans Isolation using a Combination of Purified Collagenase and Neutral Protease
Published on: September 7, 2012
Comparison of Neutral Proteases and Collagenase Class I as Essential Enzymes for Human Islet Isolation
Heide Brandhorst1, Manfred Kurfürst2, Paul R Johnson3
1Nuffield Department of Surgical Sciences, University of Oxford, Oxford, United Kingdom.; Oxford Centre for Diabetes, Endocrinology and Metabolism, Oxford, United Kingdom.; Department of Immunology, Genetics and Pathology, University of Uppsala, Uppsala, Sweden.
Unlabelled:
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.
