Abstract: Background Stem cells have been documented as a new therapeutic method for ovarian injuries such as premature ovarian failure (POF). However, effects of exosomes (Exos) derived from human endometrial stem cells (EnSCs) on diminished ovarian failure remain to be carefully elucidated. Therefore, our study aims to investigate the mechanisms of EnSC-Exos is related to Hippo signal pathway in the recovery of the cisplatin-induced granulosa cell injury model in vitro and the POF mice model in vivo. Methods Exosomes derived from human EnSCs were isolated by ultracentrifugation and identified by electron microscopy and western blot (WB) analysis, stained by PKH26, respectively. The mechanism involving suppression of Hippo signaling pathway and activation of Yes-associated protein (YAP) was investigated for evaluating EnSC-Exos therapeutic effects in the cisplatin-induced granulosa cell injury model in vitro using flow cytometry, WB, qRT-PCR, immunofluorescence and EdU staining. Hematoxylin-eosin staining, ELISA and immunohistochemistry analysis were used for evaluating EnSC-Exos therapeutic effects in POF mice model. Results We established successful construction of the cisplatin-induced granulosa cell injury model and evaluated Hippo signaling pathway activation in cisplatin-damaged granulosa cells (GCs). Furthermore, laser scanning confocal microscope and immunofluorescence demonstrated that EnSC-Exos can be transferred to cisplatin-damaged GCs to decrease apoptosis. In addition, the enhanced expression of YAP at the protein level as well as YAP/TEAD target genes, such as CTGF, ANKRD1, and the increase of YAP into the nucleus in immunofluorescence staining after the addition of EnSC-Exos to cisplatin-damaged GCs confirmed the suppression of Hippo signaling, While in vivo, EnSC-Exos successfully remedied POF in mice. Conclusions Collectively, our findings suggest that human EnSC-Exos is effective in recovery of ovarian function by chemotherapy-induced POF via activating YAP and inhibiting the Hippo signaling pathway. These findings provide new insights for further understanding of EnSC-Exos in the recovery of ovary function.

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