Comparison of Clostripain and Neutral Protease as Supplementary Enzymes for Human Islet Isolation
| Autor: | Olle Korsgren, Johanna Mönch, H Brandhorst, Manfred Kurfürst, Daniel Brandhorst, Paul Johnson |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Male Proteases Cellbiologi Biomedical Engineering Islets of Langerhans Transplantation lcsh:Medicine Endocrinology and Diabetes human islet transplantation Andrology 03 medical and health sciences Islets of Langerhans 0302 clinical medicine Equivalent medicine Humans Collagenases Aged chemistry.chemical_classification Transplantation Clostripain geography geography.geographical_feature_category clostripain lcsh:R Original Articles Cell Biology Middle Aged Islet collagenase Cysteine Endopeptidases 030104 developmental biology Enzyme chemistry neutral protease Endokrinologi och diabetes Collagenase Female Digestion 030217 neurology & neurosurgery human islet isolation medicine.drug Peptide Hydrolases |
| Zdroj: | Cell Transplantation, Vol 28 (2019) Cell Transplantation |
| ISSN: | 1555-3892 0963-6897 |
| Popis: | Although human islet transplantation has been established as valid and safe treatment for patients with type 1 diabetes, the utilization rates of human pancreases for clinical islet transplantation are still limited and substantially determined by the quality and composition of collagenase blends. While function and integrity of collagenase has been extensively investigated, information is still lacking about the most suitable supplementary neutral proteases. The present study compared islet isolation outcome after pancreas digestion by means of collagenase used alone or supplemented with either neutral protease (NP), clostripain (CP), or both proteases. Decent amounts of islet equivalents (IEQ) were isolated using collagenase alone (3090 +/- 550 IEQ/g), or in combination with NP (2340 +/- 450 IEQ/g) or CP (2740 +/- 280 IEQ/g). Nevertheless, the proportion of undigested tissue was higher after using collagenase alone (21.1 +/- 1.1%, P < 0.05) compared with addition of NP (13.3 +/- 2.2%) or CP plus NP (13.7 +/- 2.6%). Likewise, the percentage of embedded islets was highest using collagenase only (13 +/- 2%) and lowest adding NP plus CP (4 +/- 1%, P < 0.01). The latter combination resulted in lowest post-culture overall survival (42.7 +/- 3.9%), while highest survival was observed after supplementation with CP (74.5 +/- 4.8%, P < 0.01). An insulin response toward glucose challenge was present in all experimental groups, but the stimulation index was significantly decreased using collagenase plus NP (2.0 +/- 0.12) compared with supplementation with CP (3.16 +/- 0.4, P < 0.001). This study demonstrates for the first time that it is possible to isolate significant numbers of human islets combining collagenase only with CP. The supplementation with CP is an effective means to substantially reduce NP activity, which significantly decreases survival and viability after culture. This will facilitate the manufacturing of enzyme blends with less harmful characteristics. |
| Databáze: | OpenAIRE |
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