Schwann Cell-Derived Exosomes Induce the Differentiation of Human Adipose-Derived Stem Cells Into Schwann Cells

Nan Zhou; Zhao Xu; Xiang Li; Sen Ren; Jing Chen; Hewei Xiong; Cheng Wang; Jiahe Guo; Yu Kang; Zhenbing Chen; Wenqing Li; Xiaofan Yang; Xing Zhang; Xiang Xu

doi:10.3389/fmolb.2021.835135

Schwann Cell-Derived Exosomes Induce the Differentiation of Human Adipose-Derived Stem Cells Into Schwann Cells

Front Mol Biosci. 2022 Jan 31:8:835135. doi: 10.3389/fmolb.2021.835135. eCollection 2021.

Authors

Nan Zhou^{1

2}, Zhao Xu¹, Xiang Li³, Sen Ren¹, Jing Chen¹, Hewei Xiong¹, Cheng Wang¹, Jiahe Guo¹, Yu Kang¹, Zhenbing Chen¹, Wenqing Li⁴, Xiaofan Yang¹, Xing Zhang², Xiang Xu¹

Affiliations

¹ Department of Hand Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China.
² Department of Orthopedics, Trauma and Reconstructive Surgery, RWTH Aachen University Hospital, Aachen, Germany.
³ Department of Clinical Laboratory, Huai'an Second People's Hospital, The Affiliated Huai'an Hospital of Xuzhou Medical University, Huai'an, China.
⁴ Department of Hand and Foot Surgery, Huazhong University of Science and Technology Union Shenzhen Hospital, Shenzhen, China.

Abstract

Adipose-derived stem cells (ADSCs) can differentiate into Schwann cells (SCs) at the site of nerve injury, where Schwann cell-derived exosomes (SC-Exos) are suspected to exert an induction effect. Our study aimed to induce the differentiation of ADSCs in vitro using SC-Exos and to investigate the mechanisms involved through miRNA sequencing. Subcutaneous fat was used to extract ADSCs. Exosomes were extracted from Schwann cell lines (RSC96) using ultracentrifugation and were able to be taken up by human ADSCs. After 8 days of induction of ADSCs by SC-Exos, phenotypic characteristics were observed by examining the expression of SC markers (S100ß, NGFR, MPZ, GFAP) through RT-qPCR, Western blot and immunofluorescence. The RNA and protein expression levels of S100ß, NGFR, MPZ, and GFAP were found to be significantly higher in the SC-Exo induction group than in the uninduced group, which was also consistent with the immunofluorescence results. Additionally, miRNA sequencing was performed on exosome-induced ADSCs, followed by bioinformatic analysis and validation of the results. According to the sequencing results, there were a total of 94 differentially expressed miRNAs. Bioinformatics analysis indicated that 3506 Gene Ontology terms and 98 Kyoto Encyclopedia of Genes and Genomes pathways were significantly enriched. Ten miRNAs, 5 target mRNAs and elevated expression of the PIK3CD/Akt pathway were validated by RT-qPCR or Western blot, which is consistent with the sequencing results. Our study demonstrates that the utility of SC-Exos is effective in inducing the differentiation of ADSCs into SCs, in which these validated differentially expressed miRNAs exert a vital effect. This work provides a new paradigm via rationally applying Schwann cell-derived exosomes as a promising therapeutic option for repairing peripheral nerve injury.

Keywords: Schwann cell; adipose-derived stem cell; differentiation; exosome; miRNA sequencing.