Xiao-Hui Tan1,2,3, Sheng-Ji Gu1,2,3, Wen-Jie Tian4,5, Wen-Peng Song2,3,6, Yang-Yang Gu2,3,7, Yi-Ming Yuan1,2,3, Xue-Song Li1,2,3, Zhong-Cheng Xin1,2,3, Sae Woong Kim4,5, Rui-Li Guan8,9,10, Woong Jin Bae11,12. 1. Department of Urology, Peking University First Hospital, Beijing, 100034, People's Republic of China. 2. Institute of Urology, Peking University, Beijing, 100034, People's Republic of China. 3. Beijing Key Laboratory of Urogenital Diseases (Male) Molecular Diagnosis and Treatment Center, Beijing, 100034, People's Republic of China. 4. Department of Urology, College of Medicine, The Catholic University of Korea, Seoul, 06591, Republic of Korea. 5. Catholic Integrative Medicine Research Institute, The Catholic University of Korea, Seoul, 06591, Republic of Korea. 6. Department of Dental Implant Center, Beijing Stomatological Hospital, School of Stomatology, Capital Medical University, Beijing, 100050, People's Republic of China. 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. 8. Department of Urology, Peking University First Hospital, Beijing, 100034, People's Republic of China. guanruili@bjmu.edu.cn. 9. Institute of Urology, Peking University, Beijing, 100034, People's Republic of China. guanruili@bjmu.edu.cn. 10. Beijing Key Laboratory of Urogenital Diseases (Male) Molecular Diagnosis and Treatment Center, Beijing, 100034, People's Republic of China. guanruili@bjmu.edu.cn. 11. Department of Urology, College of Medicine, The Catholic University of Korea, Seoul, 06591, Republic of Korea. bwoong@catholic.ac.kr. 12. Catholic Integrative Medicine Research Institute, The Catholic University of Korea, Seoul, 06591, Republic of Korea. bwoong@catholic.ac.kr.
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.
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.