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Extracellular vesicles derived from human Sertoli cells: characterizations, proteomic analysis, and miRNA profiling

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Abstract

Background

Extracellular vesicles (EVs) contain thousands of proteins and nucleic acids, playing an important role in cell–cell communications. Sertoli cells have been essential in the testis as a “nurse cell”. However, EVs derived from human Sertoli cells (HSerCs) have not been well investigated.

Methods

EVs were isolated from HSerCs via ultracentrifugation and characterized by transmission electron microscopy, tunable resistive pulse sensing, and Western blotting. The cargo carried by HSerCs-EVs was measured via liquid chromatography-mass spectrometry and GeneChip miRNA Arrays. Bioinformatic analysis was performed to reveal potential functions of HSerCs-EVs.

Results

A total of 860 proteins with no less than 2 unique peptides and 88 microRNAs with high signal values were identified in HSerCs-EVs. Biological processes related to molecular binding, enzyme activity, and regulation of cell cycle were significantly enriched. Specifically, many proteins in HSerCs-EVs were associated with spermatogenesis and regulation of immune system, including Septins, Large proline-rich protein BAG6, Clusterin, and Galectin-1. Moreover, abundant microRNAs within HSerCs-EVs (miR-638, miR-149-3p, miR-1246, etc.) had a possible impact on male reproductive disorders such as asthenozoospermia and oligozoospermia.

Conclusions

Our study has shown that HSerCs-EVs contain diverse components such as proteins and microRNAs. Further research is required to evaluate HSerCs-EVs in spermatogenesis, which are underutilized but highly potent resources with particular promise for male infertility.

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Abbreviations

EVs:

Extracellular vesicles

SCs:

Sertoli cells

HSerCs:

Human Sertoli cells

LC:

Liquid chromatography

miRNA:

MicroRNA

MS:

Mass spectrometry

PBS:

Phosphate-buffered saline

GO:

Gene ontology

KEGG:

Kyoto encyclopedia of genes and genomes

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Acknowledgements

The authors would like to thank the donor of human Sertoli cells, who made this study possible. We also appreciate the Human Protein Atlas and the PRIDE, GEO, GO, KEGG databases for providing their platforms as well as contributors for uploading their meaningful datasets.

Funding

The work was supported by the Horizontal Projects and the National Natural Science Foundation of China (Grant No. 81401194).

Author information

Author notes

  1. Xiao-Hui Tan and Sheng-Ji Gu have contributed equally to this work.

Authors and Affiliations

  1. Department of Urology, Peking University First Hospital, Beijing, 100034, People’s Republic of China

    Xiao-Hui Tan, Sheng-Ji Gu, Yi-Ming Yuan, Xue-Song Li, Zhong-Cheng Xin & Rui-Li Guan

  2. Institute of Urology, Peking University, Beijing, 100034, People’s Republic of China

    Xiao-Hui Tan, Sheng-Ji Gu, Wen-Peng Song, Yang-Yang Gu, Yi-Ming Yuan, Xue-Song Li, Zhong-Cheng Xin & Rui-Li Guan

  3. Beijing Key Laboratory of Urogenital Diseases (Male) Molecular Diagnosis and Treatment Center, Beijing, 100034, People’s Republic of China

    Xiao-Hui Tan, Sheng-Ji Gu, Wen-Peng Song, Yang-Yang Gu, Yi-Ming Yuan, Xue-Song Li, Zhong-Cheng Xin & Rui-Li Guan

  4. Department of Urology, College of Medicine, The Catholic University of Korea, Seoul, 06591, Republic of Korea

    Wen-Jie Tian, Sae Woong Kim & Woong Jin Bae

  5. Catholic Integrative Medicine Research Institute, The Catholic University of Korea, Seoul, 06591, Republic of Korea

    Wen-Jie Tian, Sae Woong Kim & Woong Jin Bae

  6. Department of Dental Implant Center, Beijing Stomatological Hospital, School of Stomatology, Capital Medical University, Beijing, 100050, People’s Republic of China

    Wen-Peng Song

  7. Department of Radiation Medicine, Institute of Systems Biomedicine, School of Basic Medical Sciences, Peking University Health Science Center, Beijing, 100191, People’s Republic of China

    Yang-Yang Gu

Authors
  1. Xiao-Hui Tan
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Contributions

Conception and design: R-LG, Y-MY. Administrative support: X-SL, Z-CX, SWK, WJB. Collection and assembly of data: X-HT, S-JG, W-JT, W-PS, Y-YG. Data analysis and interpretation: X-HT, S-JG. Manuscript writing: X-HT, S-JG. Final approval of manuscript: All authors.

Corresponding authors

Correspondence to Rui-Li Guan or Woong Jin Bae.

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Conflicts of interest

The authors declare that they have no conflicts of interest.

Ethical approval

Ethical approval was waived since this article does not contain any studies with human participants or animals. Human Sertoli cells were commercially available and purchased from ScienCell Research Laboratories (Cat. No. 4520; Carlsbad, CA, USA).

Data availability

All data generated or analyzed during this study are included in this published article and its supplementary information files, which are available on reasonable request. The microarray data have been available in the Gene Expression Omnibus (GEO) database (http://www.ncbi.nlm.nih.gov/geo/) under the accession number GSE192696. Raw mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium via the PRIDE partner repository with the dataset identifier PXD030640.

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Tan, XH., Gu, SJ., Tian, WJ. et al. Extracellular vesicles derived from human Sertoli cells: characterizations, proteomic analysis, and miRNA profiling. Mol Biol Rep 49, 4673–4681 (2022). https://doi.org/10.1007/s11033-022-07316-1

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  • DOI: https://doi.org/10.1007/s11033-022-07316-1

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