Department of Veterans Affairs Tennessee Valley

2 ARTICLES PUBLISHED IN JoVE

Developmental Biology

Analysis of Non-Human Primate Pancreatic Islet Oxygen Consumption

Joseph M. Elsakr¹, Charles Deeter², Valerie Ricciardi³, Maureen Gannon^1,4,5,6

¹Department of Molecular Physiology and Biophysics, Vanderbilt University, ²Agilent Technologies, ³Department of Biological Sciences, Vanderbilt University, ⁴Department of Veterans Affairs Tennessee Valley, ⁵Department of Medicine, Vanderbilt University Medical Center, ⁶Department of Cell and Developmental Biology, Vanderbilt University

This protocol demonstrates the accurate and reproducible measurement of oxygen consumption in non-human primate pancreatic islets. The islet loading techniques and coating of the microplate provide a framework for efficient measurement of respiration in other types of cultured spheroids.

Preparation of Frozen Non-Human Primate Fetal Islets for Combined Single Nuclei RNA-Sequencing and ATAC-Sequencing, and Bulk Metabolomics

Darian T. Carroll¹, Sarah R. Lindsley², Melissa Kirigiti², Alex Buko³, Angela Jones⁴, Paul Kievit², Maureen Gannon^1,5,6,7

¹Department of Molecular Physiology and Biophysics, Vanderbilt University, ²Division of Metabolic Health and Disease, Oregon National Primate Research Center, ³Human Metabolome Technologies (HMT) America, ⁴Vanderbilt Technologies for Advanced Genomics, Vanderbilt University Medical Center, ⁵Department of Medicine, Vanderbilt University Medical Center, ⁶Department of Veterans Affairs Tennessee Valley, ⁷Department of Cell and Developmental Biology, Vanderbilt University

This protocol describes procedures to isolate high-quality nuclei from frozen non-human primate pancreatic islets for use in single-nucleus simultaneous RNA sequencing and ATAC sequencing while preserving the bulk cytosolic fraction for metabolomic analyses from the same samples.