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Optimizing human endometrial mesenchymal stem cells for maximal induction of angiogenesis

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Abstract

Human endometrial mesenchymal stem cells (hEMSCs) have been shown to promote neo-vascularization; however, its angiogenic function lessens with age. To determine the optimal conditions for maximizing hEMSC angiogenic capacity, we examined the effects of serial passaging on hEMSC activity. hEMSCs were cultured from passages (P) 3, 6, 9, and 12, and analyzed for proliferation, migration, differentiation and senescence, as well as their capacity to induce angiogenesis. The results showed that hEMSC proliferation and migration significantly decreased after P12. Furthermore, hEMSC differentiation into adipogenic and osteogenic lineages, as well as their proangiogenic capacity, gradually decreased from P9-12, while senescence only occurred after P12. Evaluation of angiogenic-related protein levels showed that both transforming growth factor β2 and Tie-2 was significantly reduced in hEMSCs at P12, compared to P3, possibly serving as the basis behind their lowered angiogenic capacity. Furthermore, in vivo angiogenesis evaluation with Matrigel plug assay showed that the optimal hEMSC to HUVEC ratio, for maximizing vessel formation, was 1:4. This study showed that hEMSC passaging was associated with lowered cellular functioning, bringing them closer to a senescent phenotype, especially after P12, thereby defining the optimal time period for cultivating fully functional hEMSCs for therapeutic applications.

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Data availability

The data presented in this study are available on request from the corresponding author.

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Funding

This work was supported by a grant from the National Natural Science Foundation of China (81702239), Key R&D program of Shanxi Province (International cooperation, 201903D421023), Shanxi Scholarship Council of China (2021–078), the Natural Science Foundation for Young Scientists of Shanxi Province (201901D211533, 201901D211324).

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Authors and Affiliations

  1. Department of Biochemistry and Molecular Biology, The Laboratory of Stem Cell Regenerative Medicine Research, Shanxi Key Laboratory of Birth, Defect and Cell Regeneration, Shanxi Medical University, No. 56, Xinjian South Road, Yingze District, Taiyuan, 030001, Shanxi, China

    Jie Zhang, Huifang Song, Xuemei Fan, Sheng He, Wenjuan Yin, Zexu Peng, Kun Yang, Hui Gong & Jun Xie

  2. Department of Anatomy, Shanxi Medical University, Taiyuan, 030001, Shanxi, China

    Huifang Song

  3. Department of Endocrinology, Shanxi Bethune Hospital, Shanxi Academy of Medical Sciences, The Third Hospital of Shanxi Medical University, Taiyuan, 030001, Shanxi, China

    Xuemei Fan & Kun Yang

  4. Department of Radiology, The First Hospital of Shanxi Medical University, Taiyuan, 030001, Shanxi, China

    Sheng He

  5. Department of Anatomy, Shanxi University of Chinese Medicine, Yuci, Taiyuan, 030001, Shanxi, China

    Xiaoyan Zhai

  6. Xi’an International Medical Center Plastic Surgery Hospital, Xi’an, 710068, Shaanxi, China

    Zhijun Wang

  7. Department of Gynecology and Obstetrics, The Second Hospital of Shanxi Medical, University, Taiyuan, 030001, Shanxi, China

    Yi Ping

  8. Department of Gynecology and Obstetrics, The First Hospital of Shanxi Medical University, Taiyuan, 030001, Shanxi, China

    Sanyuan Zhang

  9. Toronto General Hospital Research Institute, University Health Network, 101 College Street, Toronto, ON, M5G 1L7, Canada

    Ren-Ke Li

Authors
  1. Jie Zhang
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  2. Huifang Song
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  3. Xuemei Fan
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  12. Sanyuan Zhang
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Contributions

JZ performed the experiments, analyzed the data, and wrote the manuscript. HS, XF, SH, WY, ZP, XZ, KY, HG, ZW, YP and SZ assisted with data acquisition and analysis. RL and JX conceived and designed the experiments. All authors have read and approved the manuscript.

Corresponding authors

Correspondence to Ren-Ke Li or Jun Xie.

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Conflicts of interest

The authors declare no conflict of interest.

Ethical approval

The study was conducted in accordance with the Declaration of Helsinki, and the protocol was approved by the Research Ethics Board (REB#: 2018026) of Shanxi Medical University and the Hospital's Ethics Committee.

Consent to human and animal participate

All animal experiments were approved by the institutional animal use and care committee of Shanxi Medical University, and carried out in accordance with the Guide for the Care and Use of Laboratory Animals (NIH, 8th Edition, 2011).

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All subjects gave their written informed consent for inclusion prior to participation in the study.

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Zhang, J., Song, H., Fan, X. et al. Optimizing human endometrial mesenchymal stem cells for maximal induction of angiogenesis. Mol Cell Biochem 478, 1191–1204 (2023). https://doi.org/10.1007/s11010-022-04572-4

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