@article{69a0efe031ea4aa0a5c497b3d956cacb,
title = "Chemically-defined generation of human hemogenic endothelium and definitive hematopoietic progenitor cells",
abstract = "Human 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.",
author = "Yun Chang and Ramizah Syahirah and Oprescu, {Stephanie N.} and Xuepeng Wang and Juhyung Jung and Cooper, {Scott H.} and Sandra Torregrosa-Allen and Elzey, {Bennett D.} and Hsu, {Alan Y.} and Randolph, {Lauren N.} and Yufei Sun and Shihuan Kuang and Broxmeyer, {Hal E.} and Qing Deng and Xiaojun Lian and Xiaoping Bao",
note = "Funding Information: LT-HSCs [50] and hPSC-derived AGM-like cells [10] are able to home to bone marrow after tail vein injection. In support of this, purified hPSC-derived mCherry+ CD45+ hematopoietic cells were injected in the duct of Cuvier of 48–52 h post-fertilization (hpf) zebrafish (Supplementary Fig. 9D), and mCherry+ cells were detected in the caudal hematopoietic tissue (CHT), a site of definitive hematopoiesis, within 1-h post-transplantation (hpt) (Supplementary Fig. 9E). Compared to control neurons, more hematopoietic cells remained in the CHT at 5 hpt (Supplementary Fig. 9F and G). We also injected the mCherry+ CD45+ hematopoietic cells directly into the circulation of c-myb knockout bloodless zebrafish embryos (Supplementary Fig. 9H) at 48–52 hpf, and mCherry+ hematopoietic cells were observed in the CHT up to 96 hpf (Supplementary Fig. 9I). Similar to cord blood CD34+ hematopoietic stem cells (CB–HSCs), hPSC-derived hematopoietic cells significantly reduced the death of bloodless zebrafish up to 4 days after transplantation (Supplementary Fig. 9J), highlighting their functional capacity. Collectively, our results demonstrated the multipotency and function of hPSC-derived definitive hematopoietic cells both in vitro and in vivo.We thank members of the Deng, Lian and Bao laboratories for technical assistance and critical reading of the manuscript, Tianxiao Han, Dr. Sean P. Palecek and Dr. David V. Schaffer for valuable discussion and advice. We also gratefully acknowledge the Purdue Flow Cytometry and Cell Separation Facility, Purdue Genomics Core Facility and the Center for Medical Genomics at Indiana University School of Medicine. This study was supported by startup funding from the Davidson School of Chemical Engineering and the College of Engineering at Purdue (X.B.), Showalter Research Trust (Young Investigator Award to X.B.), NIH NIAMS (grant no. F31AR077424 to S.N.O. R01AR078695 to S.K.), NIH NCI (grant no. R01CA212609 to S.K.), NIH NIGMS (grant no. R35GM119787 to Q.D.), NIH NHLBI (grant no. R35HL139599 to H.E.B.), and NIH NIDDK (grant no. U54DK106846). Funding Information: We thank members of the Deng, Lian and Bao laboratories for technical assistance and critical reading of the manuscript, Tianxiao Han, Dr. Sean P. Palecek and Dr. David V. Schaffer for valuable discussion and advice. We also gratefully acknowledge the Purdue Flow Cytometry and Cell Separation Facility, Purdue Genomics Core Facility and the Center for Medical Genomics at Indiana University School of Medicine . This study was supported by startup funding from the Davidson School of Chemical Engineering and the College of Engineering at Purdue (X.B.), Showalter Research Trust (Young Investigator Award to X.B.), NIH NIAMS (grant no. F31AR077424 to S.N.O., R01AR078695 to S.K.), NIH NCI (grant no. R01CA212609 to S.K.), NIH NIGMS (grant no. R35GM119787 to Q.D.), NIH NHLBI (grant no. R35HL139599 to H.E.B.), and NIH NIDDK (grant no. U54DK106846 ). Publisher Copyright: {\textcopyright} 2022",
year = "2022",
month = jun,
doi = "10.1016/j.biomaterials.2022.121569",
language = "English (US)",
volume = "285",
journal = "Biomaterials",
issn = "0142-9612",
publisher = "Elsevier BV",
}
