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Clostripain, the Missing Link in the Enzyme Blend for Efficient Human Islet Isolation.


ABSTRACT:

Unlabelled

Effective digestive enzymes are crucial for successful islet isolation. Supplemental proteases are essential as they synergize with collagenase for effective pancreas digestion. The presence of tryptic-like activity has been implicated in efficient enzyme blends and the present study aimed to evaluate if addition of clostripain, an enzyme with tryptic-like activity, could improve efficacy of the islet isolation procedure.

Methods

Clostripain was added to the enzyme blend just before pancreas perfusion. Islets were isolated per standard method and numerous isolation parameters, islet quality control, and the number of isolations fulfilling standard transplantation criteria were evaluated. Two control organs per clostripain organ were chosen by blindly matching against body mass index, cold ischemia time, hemoglobin A1c, donor sex, and donor age.

Results

There were no differences in pancreas weight, dissection time, digestion time, harvest time, percent digested pancreas, or total pellet volume before islet purification between control or clostripain pancreases. Glucose-stimulated insulin release results were similar between groups. Total isolation islet equivalents, purified tissue volume and islet equivalents/g pancreas as well as fulfillment of transplantation criteria favored clostripain processed pancreases.

Conclusions

The addition of clostripain to the enzyme blend soundly improved islet yields and transplantation rates. It gently aided pancreas digestion and maintained proper islet functionality. The addition of clostripain to the enzyme blend has now been implemented into standard isolation protocols at the isolation centers in Uppsala and in Oslo.

SUBMITTER: Stahle M 

PROVIDER: S-EPMC4946465 | biostudies-literature | 2015 Jun

REPOSITORIES: biostudies-literature

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Clostripain, the Missing Link in the Enzyme Blend for Efficient Human Islet Isolation.

Ståhle Magnus M   Foss Aksel A   Gustafsson Bengt B   Lempinen Marko M   Lundgren Torbjörn T   Rafael Ehab E   Tufveson Gunnar G   Korsgren Olle O   Friberg Andrew A  

Transplantation direct 20150624 5


<h4>Unlabelled</h4>Effective digestive enzymes are crucial for successful islet isolation. Supplemental proteases are essential as they synergize with collagenase for effective pancreas digestion. The presence of tryptic-like activity has been implicated in efficient enzyme blends and the present study aimed to evaluate if addition of clostripain, an enzyme with tryptic-like activity, could improve efficacy of the islet isolation procedure.<h4>Methods</h4>Clostripain was added to the enzyme blen  ...[more]

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