Abstract
In recent years, the discovery of ovarian germ stem cells (OGSCs) has provided a new research direction for the treatment of female infertility. The ovarian microenvironment affects the proliferation and differentiation of OGSCs, and immune cells and related cytokines are important components of the microenvironment. However, whether improving the ovarian microenvironment can regulate the proliferation of OGSCs and remodel ovarian function has not been reported. In this study, we chelated chito-oligosaccharide (COS) with fluorescein isothiocyanate (FITC) to track the distribution of COS in the body. COS was given to mice through the best route of administration, and the changes in ovarian and immune function were detected using assays of organ index, follicle counting, serum estrogen (E2) and anti-Mullerian hormone (AMH) levels, and the expression of IL-2 and TNF-α in the ovaries. We found that COS significantly increased the organ index of the ovary and immune organs, reduced the rate of follicular atresia, increased the levels of E2 and AMH hormones, and increased the protein expression of IL-2 and TNF-α in the ovary. Then, COS and OGSCs were co-cultured to observe the combination of COS and OGSCs, and measure the survival rate of OGSCs. With increasing time, the fluorescence intensity of cells gradually increased, and the cytokines IL-2 and TNF-α significantly promoted the proliferation of OGSCs. In conclusion, COS could significantly improve the ovarian and immune function of chemotherapy model mice, and improve the survival rate of OGSCs, which provided a preliminary blueprint for further exploring the mechanism of COS in protecting ovarian function.
Highlights
Ovarian dysfunction includes a “natural” decline in ovarian function caused by age factors and pathological ovarian function decline caused by pathogenic factors such as radiotherapy, chemotherapy and surgery
Alkaline phosphatase (ALP) test results showed that OGSCs were weakly positive (Fig. 5c). Messenger ribonucleic acid (mRNA) of ovarian germ stem cells and the whole ovary after 7 days was extracted to detect the expression of related markers by RT-Polymerase Chain Reaction (PCR), and the results showed that OGSCs expressed MVH, fragilis, DAZL, OCT4
The results showed that OGSCs were all expressed, confirming that OGSCs were proliferative ovarian reproductive stem cells, rather than formations of multiple cells adhering to one another (Fig. 5e, f)
Summary
Ovarian dysfunction includes a “natural” decline in ovarian function caused by age factors and pathological ovarian function decline caused by pathogenic factors such as radiotherapy, chemotherapy and surgery. It was found that ovoid cells in the epithelial layer of the ovary simultaneously expressed the germ cell specific marker MVH and the proliferation of cell marker BrdU using double-immunofluorescence staining. This discovery invalidated the “fixation theory of primordial follicle pool” and suggested for the first time the hypothesis that germ stem cells exist in the ovary. When OGSC nests are destroyed, ovarian dysfunction occurs [15, 16]. Once the mechanism of OGSCs has been thoroughly studied, it might bring new hope for the clinical treatment of infertility patients caused by chemotherapy
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