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Human Pancreatic Islet Isolation: Part I: Digestion and Collection of Pancreatic Tissue
Published on: May 26, 2009
Islet isolation from human pancreas with extended cold ischemia time
W M Kühtreiber1, L T Ho, A Kamireddy
1Prodo Laboratories Inc, Irvine, California 92618, USA. wkuhtreiber@prodolabs.com
Transplantation Proceedings
|August 10, 2010
Summary
Optimizing islet isolation for extended cold ischemia time (CIT) yields high-quality human islets. This improved process ensures better islet recovery and function, crucial for transplantation success.
Area of Science:
- Transplantation immunology
- Islet biology
- Bioprocessing
Background:
- Extended cold ischemia time (CIT) over 8 hours typically reduces human islet yield and quality.
- Current islet isolation protocols face challenges with pancreata subjected to prolonged preservation.
Purpose of the Study:
- To optimize the human islet isolation process for pancreata with extended CIT.
- To evaluate novel cGMP-grade enzymes and culture media for improved islet recovery and function.
Main Methods:
- Processing of 16 pancreata with an average CIT of 13.2 hours.
- Utilized optimized digestion with cGMP collagenase and purification on continuous Euroficoll gradients.
- Islet culture in Prodo cGMP media or Miami 1A media, followed by glucose-stimulated insulin secretion (GSIS) assays.
Main Results:
- Achieved an average postpurification yield of 359 KIEQ with 13.5% purification loss.
- Isolated islets demonstrated high purity (79.7%) and viability (92%) after culture.
- GSIS assays showed a stimulation index of 6.8, indicating good islet function after extended culture.
Conclusions:
- The optimized human islet isolation process successfully recovers high-quality islets from pancreata with extended CIT.
- The developed cGMP islet culture media demonstrated superior performance compared to standard CMRL-based media.
