1. Efficient generation of human NOTCH ligand-expressing haemogenic endothelial cells as infrastructure for in vitro haematopoiesis and lymphopoiesis.
- Author
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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, and Stanley EG
- Subjects
- Humans, Endothelial Cells metabolism, Endothelial Cells cytology, Cell Differentiation, Cell Lineage genetics, Interleukin-7 metabolism, Interleukin-7 genetics, Pluripotent Stem Cells metabolism, Pluripotent Stem Cells cytology, Calcium-Binding Proteins metabolism, Calcium-Binding Proteins genetics, Adaptor Proteins, Signal Transducing metabolism, Adaptor Proteins, Signal Transducing genetics, Killer Cells, Natural metabolism, Killer Cells, Natural cytology, Hemangioblasts metabolism, Hemangioblasts cytology, Hematopoietic Stem Cells cytology, Hematopoietic Stem Cells metabolism, Homeodomain Proteins metabolism, Homeodomain Proteins genetics, Single-Cell Analysis methods, Receptors, Notch metabolism, Receptors, Notch genetics, Hematopoiesis genetics, Lymphopoiesis genetics
- 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., (© 2024. The Author(s).)
- Published
- 2024
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