Recovery of viable endocrine-specific cells and transcriptomes from human pancreatic islet-engrafted mice.

Autor: Redick SD; Diabetes Center of Excellence, Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA, USA., Leehy L; Diabetes Center of Excellence, Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA, USA., Rittenhouse AR; Department of Microbiology and Physiological Systems, University of Massachusetts Medical School, Worcester, MA, USA., Blodgett DM; Diabetes Center of Excellence, Department of Medicine, University of Massachusetts Medical School, Worcester, MA, USA.; Math and Science Division, Babson College, Wellesley, MA, USA., Derr AG; Diabetes Center of Excellence, Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA, USA.; Program in Bioinformatics and Integrative Biology, University of Massachusetts Medical School, Worcester, MA, USA., Kucukural A; Program in Bioinformatics and Integrative Biology, University of Massachusetts Medical School, Worcester, MA, USA., Garber MG; Program in Bioinformatics and Integrative Biology, University of Massachusetts Medical School, Worcester, MA, USA., Shultz LD; The Jackson Laboratory, Bar Harbor, ME, USA., Greiner DL; Diabetes Center of Excellence, Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA, USA., Wang JP; Diabetes Center of Excellence, Department of Medicine, University of Massachusetts Medical School, Worcester, MA, USA., Harlan DM; Diabetes Center of Excellence, Department of Medicine, University of Massachusetts Medical School, Worcester, MA, USA., Bortell R; Diabetes Center of Excellence, Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA, USA., Jurczyk A; Diabetes Center of Excellence, Program in Molecular Medicine, University of Massachusetts Medical School, Worcester, MA, USA.
Jazyk: angličtina
Zdroj: FASEB journal : official publication of the Federation of American Societies for Experimental Biology [FASEB J] 2020 Jan; Vol. 34 (1), pp. 1901-1911. Date of Electronic Publication: 2019 Dec 10.
DOI: 10.1096/fj.201901022RR
Abstrakt: Human 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 β, α, and δ 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 β, α, and δ 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 β 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.
(© 2019 The Authors. The FASEB Journal published by Wiley Periodicals, Inc. on behalf of Federation of American Societies for Experimental Biology.)
Databáze: MEDLINE
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