| The ratio between class II and class I collagenase determines the amount of neutral protease activity required for efficient islet release from the rat pancreas. | |
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MedLine Citation:
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PMID: 15808598 Owner: NLM Status: MEDLINE |
Abstract/OtherAbstract:
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Previous investigations clearly showed that the successful release of islets from the pancreas is mediated by both neutral protease (NP) and collagenase, consisting of subclasses I and II showing different capacities to cleave islets from the pancreas. Since no informations about the optimal ratio between class II and class I collagenase (II/I-ratio) are available yet, the present study sought to evaluate the efficient range for the II/I-ratio. METHODS: Following intraductal pancreas collagenase distension, rat islets were isolated utilizing 20 PZ-U Serva collagenase NB 1 and 1.0 or 0.4 DMC-U NP. After purification we determined the islet yield (IEQ), viability (trypan-blue exclusion) and function in diabetic nude mice. RESULTS: At 1.0 DMC-U NP, a II/I-ratio of 2.6, 1.5 or 0.7 yielded 2200 +/- 280, 2185 +/- 420, and 2205 +/-90 IEQ, respectively (ns). Viability varied between 70% and 80% (ns). Digestion time was significantly lowest (P < .05) using a II/I-ratio of 0.7. Utilization of 0.4 DMC-U NP resulted in a viability of >98% among all experimental groups (P < .001 vs 1.0 DMC-U). Islet yield decreased at a II/I-ratio of 2.6 (1520 +/- 120 IEQ, P < .05) and 1.5 (1780 +/- 130 IEQ, ns), but not at 0.7 (2310 +/- 160 IEQ, ns). Again, digestion time was lowest (P < .001) using a II/I- ratio of 0.7. Transplantation into diabetic nude mice demonstrated islet function in all experimental groups. CONCLUSIONS: NP significantly affects islet viability. This study indicates that the minimal amount of NP required for efficient islet cleavage depends on the II/I-ratio. |
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Authors:
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H Brandhorst; A Alt; S Huettler; N Raemsch-Guenther; M Kurfuerst; R G Bretzel; D Brandhorst |
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Publication Detail:
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Type: Journal Article; Research Support, N.I.H., Extramural; Research Support, U.S. Gov't, P.H.S. |
Journal Detail:
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Title: Transplantation proceedings Volume: 37 ISSN: 0041-1345 ISO Abbreviation: Transplant. Proc. Publication Date: 2005 Jan-Feb |
Date Detail:
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Created Date: 2005-04-05 Completed Date: 2005-10-28 Revised Date: 2007-11-14 |
Medline Journal Info:
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Nlm Unique ID: 0243532 Medline TA: Transplant Proc Country: United States |
Other Details:
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Languages: eng Pagination: 215-6 Citation Subset: IM |
Affiliation:
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Third Medical Department, University Hospital, Giessen, Germany. heide.brandhorst@innere.med.uni-giessen.de |
Export Citation:
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| MeSH Terms | |
Descriptor/Qualifier:
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Animals Cell Separation / methods Cell Survival Collagenases* Islets of Langerhans / cytology* Pancreas / cytology* Peptide Hydrolases / metabolism* Rats |
| Grant Support | |
ID/Acronym/Agency:
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DK 56962-2/DK/NIDDK NIH HHS |
| Chemical | |
Reg. No./Substance:
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EC 3.4.-/Peptide Hydrolases; EC 3.4.24.-/Collagenases |
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine
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