The importance of tryptic-like activity in purified enzyme blends for efficient islet isolation

Brandhorst, Heide,1962-Uppsala universitet,Enheten för klinisk immunologi,Olle Korsgren (author)

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.

Friberg, AndrewUppsala universitet,Enheten för klinisk immunologi,Olle Korsgren (author)
Andersson, Helena H. (author)
Felldin, Maria (author)
Foss, Aksel (author)
Salmela, Kaija (author)
Lundgren, TorbjörnKarolinska Institutet (author)
Tibell, AnnikaKarolinska Institutet (author)
Tufveson, GunnarUppsala universitet,Transplantationskirurgi(Swepub:uu)gutuf839 (author)
Korsgren, OlleUppsala universitet,Enheten för klinisk immunologi,Olle Korsgren(Swepub:uu)ollekors (author)
Brandhorst, Daniel,1961-Uppsala universitet,Enheten för klinisk immunologi,Olle Korsgren(Swepub:uu)danbr162 (author)
Uppsala universitetEnheten för klinisk immunologi (creator_code:org_t)

By the author/editor: Brandhorst, Heid ...; Friberg, Andrew; Andersson, Helen ...; Felldin, Maria; Foss, Aksel; Salmela, Kaija; show more...; Lundgren, Torbjö ...; Tibell, Annika; Tufveson, Gunnar; Korsgren, Olle; Brandhorst, Dani ...; show less...

About the subject

MEDICAL AND HEALTH SCIENCES: MEDICAL AND HEAL ...; and Clinical Medicin ...; and Surgery

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