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Prophylactic administration of human amniotic fluid stem cells suppresses inflammation-induced preterm birth via macrophage polarization

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Abstract

Ascending inflammation from the vagina is a major cause of preterm birth. Currently, this condition—especially when uncontrolled—has no effective treatment. Human amniotic fluid stem cells (hAFSCs) are mesenchymal stem cells known to exert potent anti-inflammatory effects in animal models of perinatal diseases, such as periventricular leukomalacia, myelomeningocele, and neonatal sepsis. However, hAFSC therapy for inflammation-induced preterm birth has not been tested. In order to determine the therapeutic effect of hAFSC transplantation, we employed a preterm mouse model of ascending infection; this model was constructed by administering lipopolysaccharide to pregnant mice. We investigated the preterm birth rate and evaluated the inflammation of tissues, which is related to progressive infections, such as those involving the cervix, placenta, and lavage cells, using real-time qPCR. Further, we tracked the fluorescence of fluorescently labeled hAFSCs using an in vivo imaging system, and hAFSC aggregation was evaluated using immunohistochemistry analysis. We also investigated the presence of multiple types of peritoneal macrophages via flow cytometry analysis. Finally, we performed sphere culturing and co-culturing to determine the therapeutic effects of hAFSCs, such as their anti-inflammatory effects and their potential to alter macrophage polarization. We found that hAFSC administration to the peritoneal cavity significantly reduced inflammation-induced preterm birth in the mouse model. The treatment also significantly suppressed inflammation of the placenta and cervix. Transplanted hAFSCs may have aggregated with peritoneal macrophages, switching them from an inflammatory to an anti-inflammatory type. This property has been reported in vivo previously, but here, we examined the effect in vitro. Our findings support the hypothesis that hAFSCs suppress inflammation and reduce preterm birth by switching macrophage polarity. This study is the first to demonstrate that hAFSCs are effective in the treatment and prevention of inflammation-induced preterm birth.

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Abbreviations

FBS:

Fetal bovine serum

hAFSCs:

Human amniotic fluid stem cells

LPS:

Lipopolysaccharide

MSCs:

Mesenchymal stem cells

α-MEM:

Alpha-modified Eagle minimum essential medium

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Acknowledgements

Not applicable.

Funding

This work was supported by JSPS Grant-in-Aid for Scientific Research (B) (grant number 20K08215) and (C) (grant number 21H03080).

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Author notes

  1. Yushi Abe and Daigo Ochiai authors equally contributed to this work.

Authors and Affiliations

  1. Department of Obstetrics & Gynecology, Keio University School of Medicine, 35 Shinanomachi, Shinjuku-ku, Tokyo, 160-8582, Japan

    Yushi Abe, Daigo Ochiai, Seiji Kanzaki, Yu Sato, Toshimitsu Otani, Satoru Ikenoue, Yoshifumi Kasuga & Mamoru Tanaka

  2. StemCell Institute Inc, Tokyo, Japan

    Seiji Kanzaki

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  1. Yushi Abe
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Contributions

YA and DO were involved in study conception and design, provision of study material or patients, collection and/or assembly of data, data analysis and interpretation, and manuscript writing. DO also provided administrative support. SK was involved in data collection and/or assembly, as well as data analysis and interpretation. YS and TO provided study material or patients, collected and/or assembled data, as well as analyzed and interpreted data. SI and YK provided administrative support and helped with data analysis and interpretation. MT was involved in study conception and design; administrative support; data collection, assembly, analysis, and interpretation; and manuscript writing. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Daigo Ochiai.

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SK is an employee of StemCell Institute Inc. Other authors have no conflict of interest to declare.

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All experimental protocols were approved by the Institutional Review Board of Keio University School of Medicine (no. 20140285), and informed consent was obtained from all the volunteer donors. All animal experiments were approved by the Animal Committee of Keio University (no. 18025).

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11010_2022_4512_MOESM1_ESM.pptx

Supplementary file1 (PPTX 5301 kb) Figure 1. Characteristics of hAFSCs. (a) Culture, (b) surface marker expression, and (c) differentiation potential of human amniotic fluid stem cells (hAFSCs), Supplementary Figure 2. Transvaginal administration of LPS.

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Abe, Y., Ochiai, D., Kanzaki, S. et al. Prophylactic administration of human amniotic fluid stem cells suppresses inflammation-induced preterm birth via macrophage polarization. Mol Cell Biochem 478, 363–374 (2023). https://doi.org/10.1007/s11010-022-04512-2

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