Human embryonic stem cells secrete macrophage migration inhibitory factor: A novel finding

Yanzhao Wei; Xiaohan Zheng; Ting Huang; Yuanji Zhong; Shengtong Sun; Xufang Wei; Qibing Liu; Tan Wang; Zhenqiang Zhao

doi:10.1371/journal.pone.0288281

Human embryonic stem cells secrete macrophage migration inhibitory factor: A novel finding

PLoS One. 2023 Aug 24;18(8):e0288281. doi: 10.1371/journal.pone.0288281. eCollection 2023.

Authors

Yanzhao Wei^{1

2

3}, Xiaohan Zheng^{1

3}, Ting Huang^{1

3}, Yuanji Zhong^{1

3}, Shengtong Sun^{1

3}, Xufang Wei^{1

3}, Qibing Liu⁴, Tan Wang^{1

3}, Zhenqiang Zhao^{1

3}

Affiliations

¹ Department of Neurology, First Affiliated Hospital of Hainan Medical University, Hainan, China.
² Department of Human Functioning, Department of Health Services, Logistics University of Chinese People's Armed Police Force, Tianjin, China.
³ Key Laboratory of Brain Science Research & Transformation in Tropical Environment of Hainan Province, Hainan Medical University, Hainan, China.
⁴ Department of Pharmacy, Hainan Medical University, Hainan, China.

Abstract

Macrophage migration inhibitory factor (MIF) is expressed in a variety of cells and participates in important biological mechanisms. However, few studies have reported whether MIF is expressed in human Embryonic stem cells (ESCs) and its effect on human ESCs. Two human ESCs cell lines, H1 and H9 were used. The expression of MIF and its receptors CD74, CD44, CXCR2, CXCR4 and CXCR7 were detected by an immunofluorescence assay, RT-qPCR and western blotting, respectively. The autocrine level of MIF was measured via enzyme-linked immunosorbent assay. The interaction between MIF and its main receptor was investigated by co-immunoprecipitation and confocal immunofluorescence microscopy. Finally, the effect of MIF on the proliferation and survival of human ESCs was preliminarily explored by incubating cells with exogenous MIF, MIF competitive ligand CXCL12 and MIF classic inhibitor ISO-1. We reported that MIF was highly expressed in H1 and H9 human ESCs. MIF was positively expressed in the cytoplasm, cell membrane and culture medium. Several surprising results emerge. The autosecreted concentration of MIF was 22 ng/mL, which was significantly higher than 2 ng/mL-6 ng/mL in normal human serum, and this was independent of cell culture time and cell number. Human ESCs mainly expressed the MIF receptors CXCR2 and CXCR7 rather than the classical receptor CD74. The protein receptor that interacts with MIF on human embryonic stem cells is CXCR7, and no evidence of interaction with CXCR2 was found. We found no evidence that MIF supports the proliferation and survival of human embryonic stem cells. In conclusion, we first found that MIF was highly expressed in human ESCs and at the same time highly expressed in associated receptors, suggesting that MIF mainly acts in an autocrine form in human ESCs.

Copyright: © 2023 Wei et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Publication types

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

MeSH terms

Blotting, Western
Cell Movement
Human Embryonic Stem Cells*
Humans
Macrophage Migration-Inhibitory Factors* / metabolism

Substances

Macrophage Migration-Inhibitory Factors

Grants and funding

This work was supported by a grant from National Natural Science Foundation of China (grant numbers 81860238); Hainan Provincial Natural Science Foundation of China (grant number 821RC694); Major Research and Development Project of Hainan Province of China (grant number ZDYF2018233); Scientific research projects in Colleges and Universities of Hainan Province of China (grant number HYYS2021A19); Project of Hainan Province Clinical Medical Center. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.