RT Journal Article SR Electronic T1 Mapping Human Hematopoietic Hierarchy at Single Cell Resolution by Microwell-seq JF bioRxiv FD Cold Spring Harbor Laboratory SP 127217 DO 10.1101/127217 A1 Shujing Lai A1 Yang Xu A1 Wentao Huang A1 Mengmeng Jiang A1 Haide Chen A1 Fang Ye A1 Renying Wang A1 Yunfei Qiu A1 Xinyi Jiang A1 Daosheng Huang A1 Jie Mao A1 Yanwei Li A1 Yingru Lu A1 Jin Xie A1 Qun Fang A1 Tiefeng Li A1 He Huang A1 Xiaoping Han A1 Guoji Guo YR 2017 UL http://biorxiv.org/content/early/2017/04/13/127217.abstract AB The classical hematopoietic hierarchy, which is mainly built with fluorescence-activated cell sorting (FACS) technology, proves to be inaccurate in recent studies. Single cell RNA-seq (scRNA-seq) analysis provides a solution to overcome the limit of FACS-based cell type definition system for the dissection of complex cellular hierarchy. However, large-scale scRNA-seq is constrained by the throughput and cost of traditional methods. Here, we developed Microwell-seq, a high-throughput and low-cost scRNA-seq platform using extremely simple devices. Using Microwell-seq, we constructed a single-cell resolution transcriptome atlas of human hematopoietic differentiation hierarchy by profiling more than 50,000 single cells throughout adult human hematopoietic system. We found that adult human hematopoietic stem and progenitor cell (HSPC) compartment is dominated by progenitors primed with lineage specific regulators. Our analysis revealed differentiation pathways for each cell types, through which HSPCs directly progress to lineage biased progenitors before differentiation. We propose a revised adult human hematopoietic hierarchy independent of oligopotent progenitors. Our study also demonstrates the broad applicability of Microwell-seq technology.