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Pancreatic Islet Isolation and Purification from Lewis Rats Using Enzymatic Digestion and Density-Gradient Separation
Published on: April 3, 2026
Islet purification method using large bottles effectively achieves high islet yield from pig pancreas
Masayuki Shimoda1, Hirofumi Noguchi, Yasutaka Fujita
1Baylor University Medical Center, Dallas, TX, USA.
Cell Transplantation
|July 17, 2012
Summary
A new bottle purification method significantly improves porcine islet yield for xenotransplantation. This method reduces mechanical damage, offering higher recovery rates and larger islet sizes compared to the standard COBE processor.
Area of Science:
- Transplantation Biology
- Islet Isolation Techniques
- Xenotransplantation Research
Background:
- Porcine islets are crucial for xenotransplantation, but isolation yields are often low due to islet fragility.
- The standard COBE 2991 cell processor (COBE) method causes mechanical damage via high shearing forces, reducing islet quality.
- Marginal quality pancreata (warm ischemic time > 40 min) present additional challenges for effective islet isolation.
Purpose of the Study:
- To evaluate a novel purification method using large plastic bottles as an alternative to the COBE processor.
- To assess the efficacy of bottle-based purification in improving islet yield and quality from porcine pancreata.
- To compare the mechanical stress and recovery rates of the bottle method versus the standard COBE method.
Main Methods:
- Ten porcine pancreata, considered marginal, were processed.
- Digested tissue was divided into three groups for purification: top loading bottle, bottom loading bottle, and standard COBE processor.
- Islet yield (IEQ/g), recovery rate (%), and average islet diameter (μm) were measured for each group.
Main Results:
- The top loading bottle method yielded significantly higher islet yield (8060 ± 1652 IEQ/g) compared to the COBE group (3900 ± 734 IE/g).
- Post-purification recovery rates were significantly higher with the top loading bottle method (99.3 ± 12.3%) versus the COBE group (49.5 ± 6.7%).
- Purified islets from both bottle methods were significantly larger (top: 156 ± 8 μm, bottom: 147 ± 6 μm) than those from the COBE group (119 ± 6 μm), indicating reduced shear force.
Conclusions:
- The novel top loading bottle purification method effectively enhances porcine islet yield and quality, especially from marginal pancreata.
- This bottle-based approach significantly reduces mechanical stress on islets compared to the standard COBE processor.
- The improved isolation efficacy makes porcine islets a more viable resource for xenotransplantation.
