Bone morphogenetic protein 4 induces hematopoietic stem cell development from murine hemogenic endothelial cells in culture

Mariko Tsuruda; Saori Morino-Koga; Xueyu Zhao; Shingo Usuki; Kei-Ichiro Yasunaga; Tomomasa Yokomizo; Ryuichi Nishinakamura; Toshio Suda; Minetaro Ogawa

doi:10.1016/j.stemcr.2024.10.005

Bone morphogenetic protein 4 induces hematopoietic stem cell development from murine hemogenic endothelial cells in culture

Stem Cell Reports. 2024 Nov 3:S2213-6711(24)00293-5. doi: 10.1016/j.stemcr.2024.10.005. Online ahead of print.

Authors

Mariko Tsuruda¹, Saori Morino-Koga¹, Xueyu Zhao¹, Shingo Usuki², Kei-Ichiro Yasunaga², Tomomasa Yokomizo³, Ryuichi Nishinakamura⁴, Toshio Suda⁵, Minetaro Ogawa⁶

Affiliations

¹ Department of Cell Differentiation, Institute of Molecular Embryology and Genetics, Kumamoto University, Kumamoto, Japan.
² Liaison Laboratory Research Promotion Center, Institute of Molecular Embryology and Genetics, Kumamoto University, Kumamoto, Japan.
³ International Research Center for Medical Sciences, Kumamoto University, Kumamoto, Japan; Department of Microscopic and Developmental Anatomy, Tokyo Women's Medical University, Tokyo, Japan.
⁴ Department of Kidney Development, Institute of Molecular Embryology and Genetics, Kumamoto University, Kumamoto, Japan.
⁵ International Research Center for Medical Sciences, Kumamoto University, Kumamoto, Japan; Institute of Hematology, Blood Diseases Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Tianjin, China.
⁶ Department of Cell Differentiation, Institute of Molecular Embryology and Genetics, Kumamoto University, Kumamoto, Japan. Electronic address: ogawamin@kumamoto-u.ac.jp.

PMID: 39547225
DOI: 10.1016/j.stemcr.2024.10.005

Abstract

Hematopoietic stem cells (HSCs) develop from hemogenic endothelial cells (HECs) during mouse embryogenesis. Understanding the signaling molecules required for HSC development is crucial for the in vitro derivation of HSCs. We previously induced HSCs from embryonic HECs, isolated at embryonic day 10.5 (E10.5), in serum-free culture conditions with stem cell factor, thrombopoietin, and an endothelial feeder layer. Here, we aimed to elucidate signal requirements for inducing HSCs from earlier-stage HECs. Single-cell RNA sequencing (RNA-seq) analysis detected bone morphogenetic protein (BMP) signaling activation in E9.5 HECs. Adding BMP4 to the culture conditions led to the induction of HSCs from E9.5 HECs. Furthermore, isolating BMP4 receptor-expressing HECs from E9.5 embryos enriched progenitors with HSC-forming ability. This study identified BMP4 as an essential factor promoting the differentiation of early HECs into HSCs, opening up new possibilities for the in vitro derivation of HSCs.

Keywords: bone marrow reconstitution; bone morphogenetic protein 4; endothelial cells; hematopoietic stem cells; hemogenic endothelial cells; mouse embryos; serum-free culture; single-cell RNA sequencing.