Recovery of viable endocrine-specific cells and transcriptomes from human pancreatic islet-engrafted mice
| Autor: | Manuel Garber, Rita Bortell, David M. Blodgett, Alan G. Derr, Dale L. Greiner, Sambra D. Redick, Leonard D. Shultz, Agata Jurczyk, David M. Harlan, Alper Kucukural, Jennifer P. Wang, Linda Leehy, Ann R. Rittenhouse |
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| Rok vydání: | 2019 |
| Předmět: |
0301 basic medicine
Adult Male Cell type insulin Transplantation Heterologous Islets of Langerhans Transplantation β cell Biology Biochemistry Diabetes Mellitus Experimental Transcriptome 03 medical and health sciences Islets of Langerhans Mice Young Adult 0302 clinical medicine graft recovery In vivo Insulin-Secreting Cells Genetics medicine Animals Humans RNA‐Seq Molecular Biology Research Articles geography geography.geographical_feature_category Pancreatic islets Gene Expression Profiling Graft Survival Cell sorting Islet Embryonic stem cell 3. Good health Cell biology 030104 developmental biology medicine.anatomical_structure L‐type voltage‐gated calcium channel Female Endocrine Cells 030217 neurology & neurosurgery Ex vivo Biotechnology Research Article |
| Zdroj: | The FASEB Journal |
| ISSN: | 1530-6860 |
| Popis: | 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. |
| Databáze: | OpenAIRE |
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