Pancreatic Islet Isolation and Purification from Lewis Rats Using Enzymatic Digestion and Density-Gradient...

Area of Science:

• Endocrinology
• Surgical Research
• Diabetes Mellitus Research

Background:

• Pancreatic islets are crucial for diabetes research and islet transplantation.
• Current limitations in islet isolation protocols hinder progress in diabetes research and therapeutics.
• Need for high-quality, standardized islet preparations for advanced research and clinical applications.

Purpose of the Study:

• To develop and present a high-throughput, standardized protocol for pancreatic islet isolation from Lewis rats.
• To optimize islet yield, purity, and viability for both preclinical and clinical applications.
• To compare traditional aliquot-based islet quantification with a semi-automated imaging-based method.

Main Methods:

• Surgical harvesting, enzymatic digestion, and density-gradient purification of pancreatic islets.
• Intraductal pancreas perfusion with a defined enzyme blend and pooled digestion of multiple pancreases.
• Density-based separation to maximize yield and minimize fragmentation, eliminating manual picking.

Main Results:

• A representative isolation yielded high-purity (98%) and viable (98%) islets.
• Functional assessment showed robust glucose-stimulated insulin secretion (10-fold increase).
• Comparison of quantification methods revealed significant variability in traditional aliquot-based counting versus automated analysis.

Conclusions:

• The developed protocol provides a reproducible and efficient method for generating high-purity, functionally intact islet preparations.
• Islets isolated using this protocol maintain robust function in vitro for up to seven days.
• The protocol supports flexibility for large-scale in vitro assays and in vivo transplantation studies.