Chang, YunSyahirah, RamizahOprescu, Stephanie N.Wang, XuepengJung, JuhyungCooper, Scott H.Torregrosa-Allen, SandraElzey, Bennett D.Hsu, Alan Y.Randolph, Lauren N.Sun, YufeiKuang, ShihuanBroxmeyer, Hal E.Deng, QingLian, XiaojunBao, Xiaoping2023-11-012023-11-012022Chang Y, Syahirah R, Oprescu SN, et al. Chemically-defined generation of human hemogenic endothelium and definitive hematopoietic progenitor cells. Biomaterials. 2022;285:121569. doi:10.1016/j.biomaterials.2022.121569https://hdl.handle.net/1805/36843Human hematopoietic stem cells (HSCs), which arise from aorta-gonad-mesonephros (AGM), are widely used to treat blood diseases and cancers. However, a technique for their robust generation in vitro is still missing. Here we show temporal manipulation of Wnt signaling is sufficient and essential to induce AGM-like hematopoiesis from human pluripotent stem cells. TGFβ inhibition at the stage of aorta-like SOX17+CD235a- hemogenic endothelium yielded AGM-like hematopoietic progenitors, which closely resembled primary cord blood HSCs at the transcriptional level and contained diverse lineage-primed progenitor populations via single cell RNA-sequencing analysis. Notably, the resulting definitive cells presented lymphoid and myeloid potential in vitro; and could home to a definitive hematopoietic site in zebrafish and rescue bloodless zebrafish after transplantation. Engraftment and multilineage repopulating activities were also observed in mouse recipients. Together, our work provided a chemically-defined and feeder-free culture platform for scalable generation of AGM-like hematopoietic progenitor cells, leading to enhanced production of functional blood and immune cells for various therapeutic applications.en-USPublisher PolicyChemically definedHematopoietic stem and progenitor cellsRNA sequencingStem cell differentiationTransplantationWnt signalingArticle