Single cell transcriptome profiling of mouse and hESC-derived pancreatic progenitors

Human embryonic stem cells (hESCs) are a potential unlimited source of insulin-producing β-cells for diabetes treatment. A greater understanding of how β-cells form during embryonic development will improve current hESC differentiation protocols. As β-cells are formed from NEUROG3-expressing endocrine progenitors, this study focused on characterizing the single-cell transcriptomes of mouse and hESC-derived endocrine progenitors. To do this, 7,223 E15.5 and 6,852 E18.5 single cells were isolated from Neurog3-Cre; Rosa26mT/mG embryos, allowing for enrichment of endocrine progenitors (yellow; tdTomato + EGFP) and endocrine cells (green; EGFP). From a NEUROG3-2A-eGFP CyT49 hESC reporter line (N5-5), 4,497 hESC-derived endocrine progenitor cells were sequenced. Differential expression analysis reveals enrichment of markers that are consistent with progenitor, endocrine, or novel cell-state populations. This study characterizes the single-cell transcriptomes of mouse and hESC-derived endocrine progenitors and serves as a resource (https://lynnlab.shinyapps.io/embryonic_pancreas/) for improving the formation of functional β-like cells from hESCs. This study characterizes the single-cell transcriptomes of mouse and hESC-derived endocrine progenitors

人类胚胎干细胞（human embryonic stem cells，hESCs）是糖尿病治疗领域产胰岛素β细胞（β-cells）的潜在无限来源。对胚胎发育过程中β细胞形成机制的深入解析，将有助于优化现有的hESC分化方案。由于β细胞由表达NEUROG3的内分泌祖细胞（endocrine progenitors）分化而来，本研究聚焦于对小鼠及hESC来源的内分泌祖细胞的单细胞转录组（single-cell transcriptomes）进行表征。为此，本研究从Neurog3-Cre; Rosa26mT/mG小鼠胚胎中分离得到7223个胚胎第15.5天（E15.5）的单细胞及6852个胚胎第18.5天（E18.5）的单细胞，实现了对内分泌祖细胞（黄色；tdTomato阳性+EGFP阳性）及内分泌细胞（绿色；EGFP阳性）的富集。本研究从NEUROG3-2A-eGFP CyT49 hESC报告细胞系（N5-5）中获取了4497个hESC来源的内分泌祖细胞并完成测序。差异表达分析结果显示，本研究富集得到的标记基因与祖细胞、内分泌细胞或新型细胞状态群体的特征相符。本研究完成了小鼠及hESC来源的内分泌祖细胞的单细胞转录组表征，同时为优化从hESCs诱导产生功能性β样细胞（β-like cells）的方法提供了参考资源（https://lynnlab.shinyapps.io/embryonic_pancreas/）。本研究完成了小鼠及hESC来源的内分泌祖细胞的单细胞转录组表征。