Recovery of viable endocrine-specific cells and transcriptomes from human pancreatic islet-engrafted mice
AuthorsRedick, Sambra D.
Rittenhouse, Ann R.
Blodgett, David M.
Derr, Alan G.
Shultz, Leonard D.
Greiner, Dale L.
Wang, Jennifer P.
Harlan, David M.
UMass Chan AffiliationsGarber Lab
Program in Bioinformatics and Integrative Biology
Department of Medicine
Department of Microbiology and Physiological Systems
Program in Molecular Medicine
Diabetes Center of Excellence
KeywordsL‐type voltage‐gated calcium channel
Cellular and Molecular Physiology
Endocrine System Diseases
Endocrinology, Diabetes, and Metabolism
Hormones, Hormone Substitutes, and Hormone Antagonists
Nucleic Acids, Nucleotides, and Nucleosides
Nutritional and Metabolic Diseases
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AbstractHuman pancreatic islets engrafted into immunodeficient mice serve as an important model for in vivo human diabetes studies. Following engraftment, islet function can be monitored in vivo by measuring circulating glucose and human insulin; however, it will be important to recover viable cells for more complex graft analyses. Moreover, RNA analyses of dissected grafts have not distinguished which hormone-specific cell types contribute to gene expression. We developed a method for recovering live cells suitable for fluorescence-activated cell sorting from human islets engrafted in mice. Although yields of recovered islet cells were relatively low, the ratios of bulk-sorted beta, alpha, and delta cells and their respective hormone-specific RNA-Seq transcriptomes are comparable pretransplant and posttransplant, suggesting that the cellular characteristics of islet grafts posttransplant closely mirror the original donor islets. Single-cell RNA-Seq transcriptome analysis confirms the presence of appropriate beta, alpha, and delta cell subsets. In addition, ex vivo perifusion of recovered human islet grafts demonstrated glucose-stimulated insulin secretion. Viable cells suitable for patch-clamp analysis were recovered from transplanted human embryonic stem cell-derived beta cells. Together, our functional and hormone-specific transcriptome analyses document the broad applicability of this system for longitudinal examination of human islet cells undergoing developmental/metabolic/pharmacogenetic manipulation in vivo and may facilitate the discovery of treatments for diabetes.
Redick SD, Leehy L, Rittenhouse AR, Blodgett DM, Derr AG, Kucukural A, Garber MG, Shultz LD, Greiner DL, Wang JP, Harlan DM, Bortell R, Jurczyk A. Recovery of viable endocrine-specific cells and transcriptomes from human pancreatic islet-engrafted mice. FASEB J. 2020 Jan;34(1):1901-1911. doi: 10.1096/fj.201901022RR. Epub 2019 Dec 10. PMID: 31914605; PMCID: PMC6972551. Link to article on publisher's site