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Collagenase does not persist in human islets following isolation
Sarah E Cross1, Stephen J Hughes, Anne Clark
1Islet Transplant Research Group, Nuffield Department of Surgical Sciences, University of Oxford, John Radcliffe Hospital, Oxford, UK. sarah.cross@nds.ox.ac.uk
Cell Transplantation
|April 5, 2012
Summary
Collagenase, an enzyme crucial for human islet isolation, does not remain within islets after the procedure. This finding alleviates safety concerns regarding residual collagenase in islet grafts for transplantation.
Area of Science:
- Transplantation immunology
- Islet cell biology
- Surgical research
Background:
- Optimal human islet isolation necessitates bacterial collagenase delivery to the pancreatic islet-exocrine interface.
- Previous studies indicated collagenase presence within human islets immediately post-intraductal administration, raising patient safety concerns.
Purpose of the Study:
- To investigate collagenase internalization into human islets during isolation.
- To determine if collagenase persists within islets after the isolation procedure and subsequent culture.
Main Methods:
- Samples from 14 clinical human islet isolations were collected at various time points: digest collection, University of Wisconsin solution incubation, immediately post-isolation, and after 24-hour culture.
- Samples were cryosectioned and immunolabeled for collagenase and insulin detection.
Main Results:
- No collagenase immunoreactivity was detected in isolated islets across all preparations and time points, except for one sample during digest collection.
- Collagenase was not found in islets immediately post-isolation or after 24-hour culture.
Conclusions:
- Collagenase that enters islets during intraductal administration is effectively removed during the collection phase.
- The absence of residual collagenase in isolated human islets alleviates significant safety concerns for islet transplantation.
