Human liver sinusoidal endothelial cells support the development of functional human pluripotent stem cell-derived Kupffer cells
Gregory M. Kent,
Michael H. Atkins,
Bryan Lung,
Adele Nikitina,
Ian M. Fernandes,
Jamie J. Kwan,
Tallulah S. Andrews,
Sonya A. MacParland,
Gordon M. Keller,
Blair K. Gage
Affiliations
Gregory M. Kent
McEwen Stem Cell Institute, University Health Network, Toronto, ON M5G1L7, Canada; Department of Medical Biophysics, University of Toronto, Toronto, ON M5G1L7, Canada
Michael H. Atkins
McEwen Stem Cell Institute, University Health Network, Toronto, ON M5G1L7, Canada
Bryan Lung
Department of Biochemistry, Schulich School of Medicine and Dentistry, Western University, London, ON N6G2V4, Canada; Department of Anatomy and Cell Biology, Schulich School of Medicine and Dentistry, Western University, London, ON N6A5C1, Canada
Adele Nikitina
McEwen Stem Cell Institute, University Health Network, Toronto, ON M5G1L7, Canada
Ian M. Fernandes
McEwen Stem Cell Institute, University Health Network, Toronto, ON M5G1L7, Canada
Jamie J. Kwan
McEwen Stem Cell Institute, University Health Network, Toronto, ON M5G1L7, Canada
Tallulah S. Andrews
Department of Biochemistry, Schulich School of Medicine and Dentistry, Western University, London, ON N6G2V4, Canada
Sonya A. MacParland
Ajmera Transplant Centre, Toronto General Research Institute, University Health Network, Toronto, ON M5G2C4, Canada; Department of Immunology, University of Toronto, Toronto, ON M5S1A8, Canada; Department of Laboratory Medicine and Pathobiology, University of Toronto, Toronto, ON M5S1A8, Canada
Gordon M. Keller
McEwen Stem Cell Institute, University Health Network, Toronto, ON M5G1L7, Canada; Department of Medical Biophysics, University of Toronto, Toronto, ON M5G1L7, Canada; Corresponding author
Blair K. Gage
Sprott Centre for Stem Cell Research, Ottawa Hospital Research Institute, Regenerative Medicine Program, Ottawa, ON K1H8L6, Canada; Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, ON K1H8M5, Canada; Corresponding author
Summary: In mice, the first liver-resident macrophages, known as Kupffer cells (KCs), are thought to derive from yolk sac (YS) hematopoietic progenitors that are specified prior to the emergence of the hematopoietic stem cell (HSC). To investigate human KC development, we recapitulated YS-like hematopoiesis from human pluripotent stem cells (hPSCs) and transplanted derivative macrophage progenitors into NSG mice previously humanized with hPSC-liver sinusoidal endothelial cells (LSECs). We demonstrate that hPSC-LSECs facilitate stable hPSC-YS-macrophage engraftment for at least 7 weeks. Single-cell RNA sequencing (scRNA-seq) of engrafted YS-macrophages revealed a homogeneous MARCO-expressing KC gene signature and low expression of monocyte-like macrophage genes. In contrast, human cord blood (CB)-derived macrophage progenitors generated grafts that contain multiple hematopoietic lineages in addition to KCs. Functional analyses showed that the engrafted KCs actively perform phagocytosis and erythrophagocytosis in vivo. Taken together, these findings demonstrate that it is possible to generate human KCs from hPSC-derived, YS-like progenitors.
