The importance of tryptic-like activity in purified enzyme blends for efficient islet isolation.
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 ef...
| Main Authors: | , , , , , , , , , , |
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| Format: | Journal article |
| Language: | English |
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2009
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| _version_ | 1797101129738223616 |
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| author | Brandhorst, H Friberg, A Andersson, H Felldin, M Foss, A Salmela, K Lundgren, T Tibell, A Tufveson, G Korsgren, O Brandhorst, D |
| author_facet | Brandhorst, H Friberg, A Andersson, H Felldin, M Foss, A Salmela, K Lundgren, T Tibell, A Tufveson, G Korsgren, O Brandhorst, D |
| author_sort | Brandhorst, H |
| collection | OXFORD |
| description | 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. |
| first_indexed | 2024-03-07T05:47:29Z |
| format | Journal article |
| id | oxford-uuid:e7b498d3-8ed3-4214-a7c6-60ce2b31dc93 |
| institution | University of Oxford |
| language | English |
| last_indexed | 2024-03-07T05:47:29Z |
| publishDate | 2009 |
| record_format | dspace |
| spelling | oxford-uuid:e7b498d3-8ed3-4214-a7c6-60ce2b31dc932022-03-27T10:40:53ZThe importance of tryptic-like activity in purified enzyme blends for efficient islet isolation.Journal articlehttp://purl.org/coar/resource_type/c_dcae04bcuuid:e7b498d3-8ed3-4214-a7c6-60ce2b31dc93EnglishSymplectic Elements at Oxford2009Brandhorst, HFriberg, AAndersson, HFelldin, MFoss, ASalmela, KLundgren, TTibell, ATufveson, GKorsgren, OBrandhorst, D 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. |
| spellingShingle | Brandhorst, H Friberg, A Andersson, H Felldin, M Foss, A Salmela, K Lundgren, T Tibell, A Tufveson, G Korsgren, O Brandhorst, D The importance of tryptic-like activity in purified enzyme blends for efficient islet isolation. |
| title | The importance of tryptic-like activity in purified enzyme blends for efficient islet isolation. |
| title_full | The importance of tryptic-like activity in purified enzyme blends for efficient islet isolation. |
| title_fullStr | The importance of tryptic-like activity in purified enzyme blends for efficient islet isolation. |
| title_full_unstemmed | The importance of tryptic-like activity in purified enzyme blends for efficient islet isolation. |
| title_short | The importance of tryptic-like activity in purified enzyme blends for efficient islet isolation. |
| title_sort | importance of tryptic like activity in purified enzyme blends for efficient islet isolation |
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