In vivo generation of bone marrow from embryonic stem cells in interspecies chimeras

• Published in: eLife Vol. 11
• Main Authors: Wen, Bingqiang, Wang, Guolun, Li, Enhong, Kolesnichenko, Olena A, Tu, Zhaowei, Divanovic, Senad, Kalin, Tanya V, Kalinichenko, Vladimir V
• Format: Journal Article
• Language: English
• Published: England eLife Science Publications, Ltd 30-09-2022; eLife Sciences Publications Ltd; eLife Sciences Publications, Ltd
• Subjects: Analysis; Animals; Aorta; B cells; Bioreactors; blastocyst complementation; Blood; Bone marrow; Bone Marrow - physiology; Bone Marrow Cells; CD44 antigen; CD47 Antigen; Cell differentiation; Cell surface; Chimera; Chimeras; Chondrocytes; Collagen (type I); Complementation; CXCL12 protein; CXCR4 protein; Embryo cells; Embryogenesis; Embryonic Stem Cells; Embryos; Fetuses; Flow cytometry; Fluorescence; Gene expression; Hematopoietic Stem Cell Transplantation; Hematopoietic stem cells; interspecies chimeras; irradiation; Ligands; Mice; Progenitor cells; Rats; RNA sequencing; Rodents; Stem cell transplantation; Stem cells; Stem Cells and Regenerative Medicine; Stromal cells; Surface markers; Syngeneic grafts; Transplantation; Japan; stem cells; interspecies chimeras; blastocyst complementation; rat; regenerative medicine; rats; mice; bone marrow; irradiation
• ISSN: 2050-084X; 2050-084X
• DOI: 10.7554/eLife.74018

Generation of bone marrow (BM) from embryonic stem cells (ESCs) promises to accelerate the development of future cell therapies for life-threatening disorders. However, such approach is limited by technical challenges to produce a mixture of functional BM progenitor cells able to replace all hematopoietic cell lineages. Herein, we used blastocyst complementation to simultaneously produce BM cell lineages from mouse ESCs in a rat. Based on fluorescence-activated cell sorting analysis and single-cell RNA sequencing, mouse ESCs differentiated into multiple hematopoietic and stromal cell types that were indistinguishable from normal mouse BM cells based on gene expression signatures and cell surface markers. Receptor-ligand interactions identified
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as major signaling pathways between hematopoietic progenitors and stromal cells. Multiple hematopoietic progenitors, including hematopoietic stem cells (HSCs) in mouse-rat chimeras derived more efficiently from mouse ESCs, whereas chondrocytes predominantly derived from rat cells. In the dorsal aorta and fetal liver of mouse-rat chimeras, mouse HSCs emerged and expanded faster compared to endogenous rat cells. Sequential BM transplantation of ESC-derived cells from mouse-rat chimeras rescued lethally irradiated syngeneic mice and demonstrated long-term reconstitution potential of donor HSCs. Altogether, a fully functional BM was generated from mouse ESCs using rat embryos as 'bioreactors'.