Department of Veterans Affairs Tennessee Valley
2 ARTICLES PUBLISHED IN JoVE
Developmental Biology
Analysis of Non-Human Primate Pancreatic Islet Oxygen Consumption
Joseph M. Elsakr 1, Charles Deeter 2, Valerie Ricciardi 3, 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. Carroll 1, Sarah R. Lindsley 2, Melissa Kirigiti 2, Alex Buko 3, Angela Jones 4, Paul Kievit 2, 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.
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