Summary

Macrophages are the first cells of the nascent immune system to emerge during embryonic development. In mice, embryonic macrophages infiltrate developing organs, where they differentiate symbiotically into tissue-resident macrophages (TRMs)1. However, our understanding of the origins and specialization of macrophages in human embryos is limited. Here we isolated CD45+ haematopoietic cells from human embryos at Carnegie stages 11 to 23 and subjected them to transcriptomic profiling by single-cell RNA sequencing, followed by functional characterization of a population of CD45+CD34+CD44+ yolk sac-derived myeloid-biased progenitors (YSMPs) by single-cell culture. We also mapped macrophage heterogeneity across multiple anatomical sites and identified diverse subsets, including various types of embryonic TRM (in the head, liver, lung and skin). We further traced the specification trajectories of TRMs from either yolk sac-derived primitive macrophages or YSMP-derived embryonic liver monocytes using both transcriptomic and developmental staging information, with a focus on microglia. Finally, we evaluated the molecular similarities between embryonic TRMs and their adult counterparts. Our data represent a comprehensive characterization of the spatiotemporal dynamics of early macrophage development during human embryogenesis, providing a reference for future studies of the development and function of human TRMs.

Overall design

We sorted CD45+CD235a- hematopoietic cells using flow cytometry from human embryos at multiple Carnegie stages (CS11, CS12, CS13, CS15, CS17, CS20 and CS23) and sites (yolk sac, head, liver, blood, lung and skin), and then performed a modified single cell tagged reverse transcription (STRT) protocol.

Contributors

Zhilei Bian # 1 2 3, Yandong Gong # 4, Tao Huang # 4, Christopher Z W Lee # 5 6, Lihong Bian # 7, Zhijie Bai # 4, Hui Shi # 4, Yang Zeng 8, Chen Liu 4, Jian He 4, Jie Zhou 8, Xianlong Li 4, Zongcheng Li 8, Yanli Ni 8, Chunyu Ma 7, Lei Cui 9, Rui Zhang 10 11, Jerry K Y Chan 12 13 14, Lai Guan Ng 5, Yu Lan 15 16, Florent Ginhoux 17 18 19 20, Bing Liu 21 22 23

Contact

bingliu17@yahoo.com.(Bing Liu)