Exosomes derived from human umbilical cord mesenchymal stem cells protect against cisplatin-induced ovarian granulosa cell stress and apoptosis in vitro

Liping Sun,Jianlu Wei,Xiaohui Deng,Kun Song,Li Li,Zhao Li,Shu Yao,Xuantao Su,Lan Chao,Beihua Kong,Xiuli Ju,Dong Li

doi:10.1038/s41598-017-02786-x

Abstract

Human umbilical cord mesenchymal stem cells (huMSCs) can treat primary ovarian insufficiency (POI) related to ovarian granulosa cell (OGC) apoptosis caused by cisplatin chemotherapy. Exosomes are a class of membranous vesicles with diameters of 30–200 nm that are constitutively released by eukaryotic cells. Exosomes mediate local cell-to-cell communication by transferring microRNAs and proteins. In the present study, we demonstrated the effects of exosomes derived from huMSCs (huMSC-EXOs) on a cisplatin-induced OGC model in vitro and discussed the preliminary mechanisms involved in these effects. We successfully extracted huMSC-EXOs from huMSC culture supernatant and observed the effective uptake of exosomes by cells with fluorescent staining. Using flow cytometry (with annexin-V/PI labelling), we found that huMSC-EXOs increased the number of living cells. Western blotting showed that the expression of Bcl-2 and caspase-3 were upregulated, whilst the expression of Bax, cleaved caspase-3 and cleaved PARP were downregulated to protect OGCs. These results suggest that huMSC-EXOs can be used to prevent and treat chemotherapy-induced OGC apoptosis in vitro. Therefore, this work provides insight and further evidence of stem cell function and indicates that huMSC-EXOs protect OGCs from cisplatin-induced injury in vitro.

Highlights

With the increased incidence of gynaecological cancer, clinical application of chemical treatment is prevalent
Studies have previously demonstrated that human umbilical cord mesenchymal stem cells (huMSCs) can be used to treat primary ovarian insufficiency (POI) and protect ovarian granulosa cell (OGC) from damage by cisplatin[15], but the exact mechanisms responsible for this protection are unclear
We considered whether the effects of huMSCs on OGCs occurred via a paracrine secretion mechanism

Summary

Introduction

With the increased incidence of gynaecological cancer, clinical application of chemical treatment is prevalent. Lee et al confirmed that after MSC transplantation, which can significantly increase the number of ovarian follicles and the oestradiol concentration, mice with chemotherapy-induced POI can maintain long-term fertility, suggesting that MSCs can help repair the ovary structure and improve ovarian function[13]. These effects may be closely related to the suppression of OGC apoptosis. MSCs directly migrate to the damaged ovaries and differentiate into follicular cells in the ovarian microenvironment to promote the recovery of reproductive endocrine function and inhibit OGC apoptosis[15]. Studies have shown that dendritic cell (DC)-derived exosomes may target and activate CD4 (+) T cells through the endocrine pathway to improve cardiac function after myocardial infarction[19,20,21]

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Conclusion