Efficient generation of human NOTCH ligand-expressing haemogenic endothelial cells as infrastructure for in vitro haematopoiesis and lymphopoiesis

Sun, S; Motazedian, A; Li, JY; Wijanarko, K; Zhu, JJ; Tharmarajah, K; Strumila, KA; Shkaruta, A; Nigos, LR; Schiesser, JV; Yu, Y; Neeson, PJ; Ng, ES; Elefanty, AG; Stanley, EG

Author(s): Sun, S; Motazedian, A; Li, JY; Wijanarko, K; Zhu, JJ; Tharmarajah, K; Strumila, KA; Shkaruta, A; Nigos, LR; Schiesser, JV; Yu, Y; Neeson, PJ; Ng, ES; Elefanty, AG; Stanley, EG;
Details: Publication Year 2024-09-04,Volume 15,Issue #1,Page 7698
Journal Title: Nature Communications
Publication Type: Research article
Abstract: Arterial endothelial cells (AECs) are the founder cells for intraembryonic haematopoiesis. Here, we report a method for the efficient generation of human haemogenic DLL4+ AECs from pluripotent stem cells (PSC). Time-series single-cell RNA-sequencing reveals the dynamic evolution of haematopoiesis and lymphopoiesis, generating cell types with counterparts present in early human embryos, including stages marked by the pre-haematopoietic stem cell genes MECOM/EVI1, MLLT3 and SPINK2. DLL4+ AECs robustly support lymphoid differentiation, without the requirement for exogenous NOTCH ligands. Using this system, we find IL7 acts as a morphogenic factor determining the fate choice between the T and innate lymphoid lineages and also plays a role in regulating the relative expression level of RAG1. Moreover, we document a developmental pathway by which human RAG1+ lymphoid precursors give rise to the natural killer cell lineage. Our study describes an efficient method for producing lymphoid progenitors, providing insights into their endothelial and haematopoietic ontogeny, and establishing a platform to investigate the development of the human blood system.
Publisher: Springer Nature
Keywords: Humans; *Hematopoiesis/genetics; *Lymphopoiesis/genetics; Endothelial Cells/metabolism/cytology; Cell Differentiation; Cell Lineage/genetics; Interleukin-7/metabolism/genetics; Pluripotent Stem Cells/metabolism/cytology; Calcium-Binding Proteins/metabolism/genetics; Adaptor Proteins, Signal Transducing/metabolism/genetics; Killer Cells, Natural/metabolism/cytology; Hemangioblasts/metabolism/cytology; Hematopoietic Stem Cells/cytology/metabolism; Homeodomain Proteins/metabolism/genetics; Single-Cell Analysis/methods; Receptors, Notch/metabolism/genetics
Department(s): Laboratory Research
Publisher's Version: https://doi.org/10.1038/s41467-024-51974-7
Terms of Use/Rights Notice: Refer to copyright notice on published article.

Creation Date: 2024-09-24 02:20:14

Last Modified: 2024-09-24 02:22:35