Human BM-MSC secretome enhances human granulosa cell proliferation and steroidogenesis and restores ovarian function in primary ovarian insufficiency mouse model

Hang-Soo Park,Mara Ulin,Mahmood Bilal,Sahar Esfandyari,Ayman Al-Hendy,Rishi Man Chugh,Abdeljabar El Andaloussi,Elie Hobeika,Amro Elsharoud,Natalia Garcia

doi:10.1038/s41598-021-84216-7

Abstract

Primary ovarian insufficiency (POI) is defined as the loss of ovarian function before 40 years of age. It clinically manifests as amenorrhea, infertility, and signs of estrogen insufficiency. POI is frequently induced by chemotherapy. Gonadotoxic chemotherapy reagents damage granulosa cells, which are essential for follicular function and development. Our recently published studies demonstrated that intraovarian transplantation of human mesenchymal stem cells (hMSCs) can restore fertility in a chemotherapy-induced POI mouse model. However, the regenerative mechanism underlying the hMSC effect in POI mice is not fully understood. Here, we report that the hMSC secretome increased the proliferation of human granulosa cells (HGrC1). We showed by FACS analysis that treatment of HGrC1 cells with hMSC-conditioned media (hMSC CM) stimulates cellular proliferation. We also demonstrated that the expression of steroidogenic enzymes involved in the production of estrogen, CYP19A1 and StAR, are significantly elevated in hMSC CM-treated HGrC1 cells. Our data suggest that hMSC CM stimulates granulosa cell proliferation and function, which may explain the therapeutic effect of hMSCs in our chemotherapy-induced POI animal model. Our findings indicate that the hMSC secretome may be a novel treatment approach for restoring granulosa cell and ovarian function in patients with POI.

Highlights

Primary ovarian insufficiency (POI) is defined as the loss of ovarian function before 40 years of age
Total caspase 3 was not significantly decreased in damaged cells treated with control media (Group I) or human mesenchymal stem cells (hMSCs) CM (Group II; Fig. 1b–e); cleaved caspase 3, which was increased by cyclophosphamide (1.00 ± 0.17 fold), was significantly decreased by hMSC-conditioned media (hMSC CM) treatment (0.71 ± 0.19 fold)
Our results suggest that hMSC CM can reverse markers of chemotherapy-induced apoptosis in granulosa cells

Summary

Introduction

Primary ovarian insufficiency (POI) is defined as the loss of ovarian function before 40 years of age. Our data suggest that hMSC CM stimulates granulosa cell proliferation and function, which may explain the therapeutic effect of hMSCs in our chemotherapy-induced POI animal model. We hypothesize that restoration of estrogen production, ovarian volume, and fertility potential in chemotherapy-induced POI mice following intraovarian injection of MSCs is due to the paracrine effects of factors produced by the MSCs. We hypothesize that restoration of estrogen production, ovarian volume, and fertility potential in chemotherapy-induced POI mice following intraovarian injection of MSCs is due to the paracrine effects of factors produced by the MSCs To test this hypothesis, we examined the effect of factors secreted by MSCs on the proliferation and function of a human nonluteinized granulosa cell line (HGrC1) cultured with conditioned media from MSCs (hMSC CM). Proliferation, and steroidogenesis in hMSC CM-treated HGrC1 to determine the mechanism underlying the effect of MSCs in chemotherapy-induced POI mice

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