Single-cell sequencing reveals the existence of fetal vascular endothelial stem cell-like cells in mouse liver

Fitriana N Rahmawati; Tomohiro Iba; Hisamichi Naito; Shota Shimizu; Hirotaka Konishi; Weizhen Jia; Nobuyuki Takakura

doi:10.1186/s13287-023-03460-y

Single-cell sequencing reveals the existence of fetal vascular endothelial stem cell-like cells in mouse liver

Stem Cell Res Ther. 2023 Aug 30;14(1):227. doi: 10.1186/s13287-023-03460-y.

Authors

Fitriana N Rahmawati¹, Tomohiro Iba², Hisamichi Naito², Shota Shimizu¹, Hirotaka Konishi¹, Weizhen Jia¹, Nobuyuki Takakura^{3

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Affiliations

¹ Department of Signal Transduction, Research Institute for Microbial Diseases, Osaka University, Suita, Osaka, Japan.
² Department of Physiology, School of Medicine, Kanazawa University, Kanazawa, Ishikawa, Japan.
³ Department of Signal Transduction, Research Institute for Microbial Diseases, Osaka University, Suita, Osaka, Japan. ntakaku@biken.osaka-u.ac.jp.
⁴ Immunology Frontier Research Center, Osaka University, Suita, Japan. ntakaku@biken.osaka-u.ac.jp.
⁵ Center for Infectious Disease Education and Research, Osaka University, Suita, Japan. ntakaku@biken.osaka-u.ac.jp.
⁶ Integrated Frontier Research for Medical Science Division, Institute for Open and Transdisciplinary Research Initiatives (OTRI), Osaka University, Suita, Japan. ntakaku@biken.osaka-u.ac.jp.

Abstract

Background: A resident vascular endothelial stem cell (VESC) population expressing CD157 and CD200 has been identified recently in the adult mouse. However, the origin of this population and how it develops has not been characterized, nor has it been determined whether VESC-like cells are present during the perinatal period. Here, we investigated the presence of perinatal VESC-like cells and their relationship with the adult VESC-like cell population.

Methods: We applied single-cell RNA sequencing of endothelial cells (ECs) from embryonic day (E) 14, E18, postnatal day (P) 7, P14, and week (W) 8 liver and investigated transcriptomic changes during liver EC development. We performed flow cytometry, immunofluorescence, colony formation assays, and transplantation assays to validate the presence of and to assess the function of CD157⁺ and CD200⁺ ECs in the perinatal period.

Results: We identified CD200^- expressing VESC-like cells in the perinatal period. These cells formed colonies in vitro and had high proliferative ability. The RNA velocity tool and transplantation assay results indicated that the projected fate of this population was toward adult VESC-like cells expressing CD157 and CD200 1 week after birth.

Conclusion: Our study provides a comprehensive atlas of liver EC development and documents VESC-like cell lineage commitment at single-cell resolution.

Keywords: Development; Endothelial stem cells; Heterogeneity; Mouse liver; RNA velocity; Single-cell transcriptomic sequencing; Trajectory analysis.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Adult Stem Cells*
Animals
Endothelial Progenitor Cells*
Endothelium, Vascular
Female
Fetus
Liver
Mice
Pregnancy