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The importance of t...
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Brandhorst, Heide,1962-Uppsala universitet,Enheten för klinisk immunologi,Olle Korsgren
(author)
The importance of tryptic-like activity in purified enzyme blends for efficient islet isolation
- Article/chapterEnglish2009
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Numbers
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LIBRIS-ID:oai:DiVA.org:uu-103604
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https://urn.kb.se/resolve?urn=urn:nbn:se:uu:diva-103604URI
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https://doi.org/10.1097/TP.0b013e31819499f0DOI
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http://kipublications.ki.se/Default.aspx?queryparsed=id:118300030URI
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Language:English
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Summary in:English
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Subject category:ref swepub-contenttype
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Subject category:art swepub-publicationtype
Notes
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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.
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Friberg, AndrewUppsala universitet,Enheten för klinisk immunologi,Olle Korsgren
(author)
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Andersson, Helena H.
(author)
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Felldin, Maria
(author)
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Foss, Aksel
(author)
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Salmela, Kaija
(author)
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Lundgren, TorbjörnKarolinska Institutet
(author)
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Tibell, AnnikaKarolinska Institutet
(author)
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Tufveson, GunnarUppsala universitet,Transplantationskirurgi(Swepub:uu)gutuf839
(author)
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Korsgren, OlleUppsala universitet,Enheten för klinisk immunologi,Olle Korsgren(Swepub:uu)ollekors
(author)
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Brandhorst, Daniel,1961-Uppsala universitet,Enheten för klinisk immunologi,Olle Korsgren(Swepub:uu)danbr162
(author)
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Uppsala universitetEnheten för klinisk immunologi
(creator_code:org_t)
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In:Transplantation87:3, s. 370-50041-13371534-6080
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Lundgren, Torbjö ...
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Tufveson, Gunnar
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Transplantation
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