Chemically-defined generation of human hemogenic endothelium and definitive hematopoietic progenitor cells

Yun Chang; Ramizah Syahirah; Stephanie N Oprescu; Xuepeng Wang; Juhyung Jung; Scott H Cooper; Sandra Torregrosa-Allen; Bennett D Elzey; Alan Y Hsu; Lauren N Randolph; Yufei Sun; Shihuan Kuang; Hal E Broxmeyer; Qing Deng; Xiaojun Lian; Xiaoping Bao

doi:10.1016/j.biomaterials.2022.121569

Chemically-defined generation of human hemogenic endothelium and definitive hematopoietic progenitor cells

Biomaterials. 2022 Jun:285:121569. doi: 10.1016/j.biomaterials.2022.121569. Epub 2022 May 6.

Authors

Affiliations

¹ Davidson School of Chemical Engineering, Purdue University, West Lafayette, IN, 47907, USA; Purdue University Center for Cancer Research, West Lafayette, IN, 47907, USA.
² Department of Biological Sciences, Purdue University, West Lafayette, IN, 47907, USA.
³ Purdue University Center for Cancer Research, West Lafayette, IN, 47907, USA; Department of Animal Sciences, Purdue University, West Lafayette, IN, 47907, USA.
⁴ Department of Microbiology and Immunology, Indiana University School of Medicine, Indianapolis, IN, 46202, USA.
⁵ Purdue University Center for Cancer Research, West Lafayette, IN, 47907, USA.
⁶ Purdue University Center for Cancer Research, West Lafayette, IN, 47907, USA; Department of Comparative Pathobiology, Purdue University, West Lafayette, IN, 47907, USA.
⁷ Department of Biological Sciences, Purdue University, West Lafayette, IN, 47907, USA; Department of Pathology, Harvard Medical School, Boston, MA, 02115, USA.
⁸ Departments of Biomedical Engineering, Biology, The Huck Institutes of the Life Sciences, The Pennsylvania State University, University Park, PA, 16802, USA.
⁹ Davidson School of Chemical Engineering, Purdue University, West Lafayette, IN, 47907, USA.
¹⁰ Purdue University Center for Cancer Research, West Lafayette, IN, 47907, USA; Department of Biological Sciences, Purdue University, West Lafayette, IN, 47907, USA. Electronic address: qingdeng@purdue.edu.
¹¹ Departments of Biomedical Engineering, Biology, The Huck Institutes of the Life Sciences, The Pennsylvania State University, University Park, PA, 16802, USA. Electronic address: Lian@psu.edu.
¹² Davidson School of Chemical Engineering, Purdue University, West Lafayette, IN, 47907, USA; Purdue University Center for Cancer Research, West Lafayette, IN, 47907, USA. Electronic address: bao61@purdue.edu.

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.

Keywords: Chemically defined; Hematopoietic stem and progenitor cells; RNA sequencing; Stem cell differentiation; Transplantation; Wnt signaling.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Cell Differentiation / physiology
Hemangioblasts*
Hematopoiesis / physiology
Hematopoietic Stem Cells
Humans
Mesonephros
Mice
Zebrafish

Abstract

Publication types

MeSH terms

Grants and funding