In vitro cultured H7 human embryonic stem cells (WiCell) and H7-derived downstream early mesoderm progenitors

Stem-cell differentiation to desired lineages requires navigating alternating developmental paths that often lead to unwanted cell types. Hence, comprehensive developmental roadmaps are crucial to channel stem-cell differentiation toward desired fates. To this end, here, we map bifurcating lineage choices leading from pluripotency to 12 human mesodermal lineages, including bone, muscle, and heart. We defined the extrinsic signals controlling each binary lineage decision, enabling us to logically block differentiation toward unwanted fates and rapidly steer pluripotent stem cells toward 80%–99% pure human mesodermal lineages at most branchpoints. This strategy enabled the generation of human bone and heart progenitors that could engraft in respective in vivo models. Mapping stepwise chromatin and single-cell gene expression changes in mesoderm development uncovered somite segmentation, a previously unobservable human embryonic event transiently marked by HOPX expression. Collectively, this roadmap enables navigation of mesodermal development to produce transplantable human tissue progenitors and uncover developmental processes. doi:10.1016/j.cell.2016.06.011

将干细胞定向分化为目标细胞谱系，需在多条交替出现的发育路径中进行抉择，而这些路径往往会导向非预期的细胞类型。因此，完整的发育图谱对于引导干细胞定向分化至预期命运至关重要。为此，本研究绘制了从多能干细胞出发，分化为12种人类中胚层谱系（涵盖骨、肌肉与心脏）的分支型谱系选择路径。我们明确了调控每一次二元谱系抉择的外源信号，借此可从逻辑上阻断向非预期命运的分化，并在绝大多数分支点处，快速将多能干细胞引导至纯度达80%~99%的人类中胚层谱系。该策略可生成可在对应体内模型中成功定植的人类骨与心脏祖细胞。通过解析中胚层发育过程中染色质与单细胞基因表达的动态变化图谱，我们发现了体节发生这一此前未被观测到的人类胚胎事件，该事件以HOPX的瞬时表达作为标志性特征。综上，本发育图谱可实现中胚层发育的精准调控，用以制备可移植的人类组织祖细胞，并揭示未知的发育进程。doi:10.1016/j.cell.2016.06.011

搜集汇总

背景与挑战

背景概述

该数据集聚焦于H7人类胚胎干细胞及其衍生的早期中胚层祖细胞的体外培养研究。通过整合RNA-Seq和ATAC-Seq技术，绘制了从多能性到12种人类中胚层谱系（如骨、肌肉和心脏）的发育路径，旨在指导干细胞分化为特定组织并生成可移植的祖细胞。数据集包含12个样本，涵盖单细胞和批量转录组以及染色质可及性数据，为研究人类中胚层发育提供了关键资源。