Summary

Definitive hematopoiesis generates hematopoietic stem/progenitor cells (HSPCs) that give rise to all mature blood and immune cells, but remains poorly defined in human. Here, we resolve human hematopoietic populations at the earliest hematopoiesis stage by single-cell RNA-seq. We characterize the distinct molecular profiling between early primitive and definitive hematopoiesis in both human embryonic stem cell (hESC) differentiation and early embryonic development. We identify CD44 to specifically discriminate definitive hematopoiesis and generate definitive HSPCs from hESCs. The multipotency of hESCs-derived HSPCs for various blood and immune cells is validated by single-cell clonal assay. Strikingly, these hESCs-derived HSPCs give rise to blood and lymphoid lineages in vivo. Lastly, we characterize gene-expression dynamics in definitive and primitive hematopoiesis and reveal an unreported role of ROCK-inhibition in enhancing human definitive hematopoiesis. Our study provides a prospect for understanding human early hematopoiesis and a firm basis for generating blood and immune cells for clinical purposes.

Overall design

10X Single-cell RNASeq of FACS-sorted CD43+ hematopoietic cells differentiated from H1 hESCs at day 8 (hESCs-HPCs), as well as FACS-sorted Lin-CD34+CD38- HSPCs mobilized in human peripheral blood (PB-HSPCs).

Contributors

Yanling Zhu 1 2, Tianyu Wang 1 2, Jiaming Gu 1 2 3, Ke Huang 1 2, Tian Zhang 1 2, Zhishuai Zhang 1 2 3, He Liu 1 2, Jun Tang 1 2, Yuchan Mai 1 2, Yanqi Zhang 1 2 3, Yuhang Li 1 2 3, Yashu Feng 4, Baoqiang Kang 1 2, Jinbing Li 1 2 3, Yongli Shan 1 2, Qianyu Chen 1 2, Jian Zhang 1 2, Bing Long 4, Junwei Wang 1 2, Minghui Gao 1 2, Di Zhang 1 2 3, Min Zhou 1 2, Xiaofen Zhong 1 2, Jiekai Chen 1 2, Duanqing Pei 1 2, Jinfu Nie 1 2, Bing Liu 5, Guangjin Pan 6 7 8 9

Contact

pan_guangjin@gibh.ac.cn.(Guangjin Pan)