McEwen Stem Cell Institute, University Health Network, Toronto, ON M5G1L7, Canada; Department of Medical Biophysics, University of Toronto, Toronto, ON M5G1L7, Canada.
McEwen Stem Cell Institute, University Health Network, Toronto, ON M5G1L7, Canada.
Cell Rep. 2024 Aug 27;43(8):114629. doi: 10.1016/j.celrep.2024.114629. Epub 2024 Aug 14.
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.
在小鼠中，最初的肝脏驻留巨噬细胞，即库普弗细胞（KCs），被认为来源于卵黄囊（YS）造血祖细胞，这些祖细胞在造血干细胞（HSC）出现之前就已经被指定了。为了研究人类 KC 的发育，我们从人类多能干细胞（hPSCs）中重新诱导出类似 YS 的造血，并将衍生的巨噬细胞祖细胞移植到先前用人 hPSC-肝窦内皮细胞（LSEC）人源化的 NSG 小鼠中。我们证明 hPSC-LSEC 可促进 hPSC-YS-巨噬细胞的稳定植入，至少持续 7 周。植入的 YS-巨噬细胞的单细胞 RNA 测序（scRNA-seq）显示出一种均质的 MARCO 表达的 KC 基因特征，以及单核细胞样巨噬细胞基因的低表达。相比之下，人脐带血（CB）衍生的巨噬细胞祖细胞生成的移植物除了 KC 外，还包含多个造血谱系。功能分析表明，植入的 KC 能够在体内积极进行吞噬作用和红细胞吞噬作用。综上所述，这些发现表明，有可能从 hPSC 衍生的、类似 YS 的祖细胞中产生人类 KC。
