Optimizing Porcine Islet Isolation to Markedly Reduce Enzyme Consumption Without Sacrificing Islet Yield or Function

Area of Science:

• Biotechnology
• Transplantation Research
• Endocrinology

Background:

• Human islet transplantation offers benefits for type 1 diabetes but faces limited donor availability.
• Porcine pancreas presents an abundant alternative for optimizing islet isolation techniques.
• Enhancing porcine islet isolation can increase patient access to life-saving procedures.

Purpose of the Study:

• To optimize porcine islet isolation protocols by evaluating the impact of varying enzyme concentrations and perfusion volumes.
• To assess the efficiency of enzyme retention and distribution during pancreatic perfusion and digestion.
• To reduce the cost and improve the scalability of islet isolation for research and clinical applications.

Main Methods:

• Porcine islet isolations were conducted using reduced concentrations of collagenase and neutral protease, and varied perfusion volumes.
• Enzyme retention and distribution within the pancreas were evaluated.
• Margin-marking dye was used to visualize perfusion solution infiltration into the tissue.

Main Results:

• Reducing enzyme usage by up to 67% and perfusion volume by 50% maintained successful islet isolation outcomes.
• Islet yield and tissue digestion percentages were comparable to control groups despite reduced reagent usage.
• Increased perfusion volume did not enhance tissue infiltration, as visualized by dye distribution.

Conclusions:

• Current porcine islet isolation protocols utilize excessive enzymes, increasing costs and hindering research.
• Optimized protocols can significantly decrease enzyme consumption while preserving islet yield and function.
• These findings accelerate the development of porcine islets for clinical transplantation in type 1 diabetes.