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

• Published in: The FASEB journal 2020-01, Vol.34 (1), p.1901-1911
• Main Authors: Redick, Sambra D., Leehy, Linda, Rittenhouse, Ann R., Blodgett, David M., Derr, Alan G., Kucukural, Alper, Garber, Manuel G., Shultz, Leonard D., Greiner, Dale L., Wang, Jennifer P., Harlan, David M., Bortell, Rita, Jurczyk, Agata
• Format: Article
• Language: English
• Subjects: Adult; Animals; Diabetes Mellitus, Experimental - metabolism; Diabetes Mellitus, Experimental - pathology; Endocrine Cells - metabolism; Endocrine Cells - physiology; Female; Gene Expression Profiling - methods; graft recovery; Graft Survival - physiology; Humans; insulin; Insulin - metabolism; Insulin-Secreting Cells - metabolism; Insulin-Secreting Cells - physiology; Islets of Langerhans - metabolism; Islets of Langerhans - physiology; Islets of Langerhans Transplantation - methods; L‐type voltage‐gated calcium channel; Male; Mice; RNA‐Seq; Transcriptome - physiology; Transplantation, Heterologous - methods; Young Adult; β cell
• ISSN: 0892-6638; 1530-6860
• DOI: 10.1096/fj.201901022RR

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.