Functional, metabolic and transcriptional maturation of human pancreatic islets derived from stem cells. Functional, metabolic and transcriptional maturation of human pancreatic islets derived from stem cells

Using 10x Genomics Chromium Single Cell 3’ RNAseq platform, we sequenced human stem cell derived pancreatic islet cells during different stages of development and maturation process. We optimized a protocol for pancreatic endocrine differentiation, particularly focusing on the final, extended maturation stage during which significant reorganization of the cytoarchitecture of islet-like aggregates took place correlating with a remarkable maturation of glucose-induced insulin secretion. The transcriptomic changes during the time-course were combined with detailed physiological studies including dynamic insulin secretion assays, respirometry measurements, electrophysiological profiling as well as imaging of calcium and cAMP signaling and exocytosis. Overall design: 18 scRNAseq samples were analyzed.

本研究依托10x Genomics Chromium单细胞3'转录组测序（10x Genomics Chromium Single Cell 3’ RNAseq）平台，对人干细胞诱导产生的胰腺胰岛细胞在发育与成熟进程的不同阶段开展测序。我们优化了胰腺内分泌分化的实验方案，重点聚焦于最终的延长成熟阶段：在此阶段，类胰岛聚集体的细胞构筑发生显著重构，且该重构与葡萄糖诱导胰岛素分泌功能的显著成熟密切相关。本研究将时序过程中的转录组变化与多项精细生理实验相结合，涵盖动态胰岛素分泌检测、呼吸计量分析、电生理特征分析，以及钙信号与环磷酸腺苷（cAMP）信号成像及胞吐作用观测。实验整体设计：共分析18个单细胞RNA测序（scRNAseq）样本。