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Preparation of Frozen Non-Human Primate Fetal Islets for Combined Single Nuclei RNA-Sequencing and ATAC-Sequencing, and Bulk Metabolomics
Published on: November 8, 2024
Pancreatic islet isolation variables in non-human primates (rhesus macaques)
P Andrades1, C K Asiedu, B Gansuvd
1Department of Surgery, University of Alabama at Birmingham, Birmingham, AL 35294-0019, USA. patoandrades@gmail.com
Diabetologia
|May 10, 2008
Summary
Optimizing non-human primate (NHP) islet isolation for pancreatic islet transplantation (PIT) requires careful control of variables. Key factors influencing islet yield and quality include pancreas preservation, enzyme specifics, and collection methods for improved preclinical PIT studies.
Area of Science:
- Endocrinology
- Transplantation Immunology
- Preclinical Research
Background:
- Non-human primates (NHPs) are crucial preclinical models for pancreatic islet transplantation (PIT) due to their biological similarity to humans.
- Challenges in NHP islet isolation include limited tissue availability, extended learning curves, and high costs, impacting study consistency.
- Identifying key variables is essential to improve the reliability of NHP islet isolation for PIT research.
Purpose of the Study:
- To identify critical variables influencing the quantity and quality of isolated non-human primate islets.
- To enhance the consistency and success rates of islet isolation procedures for preclinical pancreatic islet transplantation studies.
Main Methods:
- Retrospective analysis of 72 consecutive islet isolations from rhesus macaques.
- Utilized a scaled-down, semi-automated islet isolation technique.
- Evaluated 22 donor, surgical, and isolation technique variables, assessing islet yield, isolation success, and transplantation outcomes.
Main Results:
- Multivariate analysis revealed that monkey type, pancreas preservation method, enzyme lot, and enzyme volume significantly impacted islet yield.
- Enzyme lot and volume were critical for successful islet isolation.
- Transplantation success correlated with pancreas preservation, enzyme lot, endotoxin levels, and the COBE collection method.
Conclusions:
- Islet quantity and quality exhibit significant variability across isolations.
- Recommendations for future NHP islet isolation include employing bilayer preservation, using >80 ml of Liberase, collecting non-fractioned tissue via COBE, and rigorous infection monitoring.
- These optimized protocols aim to improve NHP islet isolation consistency for advancing PIT research.
