Experimental animal studies on the effect of ovarian tissue vitrification-thawed-transplantation and in vitro fertilization outcome
10.3760/cma.j.cn101441-20221212-00559
- VernacularTitle:卵巢组织玻璃化冷冻移植效果及体外受精结局的动物实验研究
- Author:
Dabing XU
1
;
Yi ZHANG
1
;
Jiayi GU
1
;
Liang WANG
1
;
Ningxia SUN
1
Author Information
1. 海军军医大学第二附属医院生殖医学中心，上海 200003
- Publication Type:Journal Article
- Keywords:
Fertilization in vitro;
Fertility preservation;
Ovarian tissue vitrification;
Ovarian tissue transplantation
- From:
Chinese Journal of Reproduction and Contraception
2023;43(2):150-157
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effects of vitrification and transplantation on mouse ovarian tissues.Methods:Female ICR mice were divided into three groups: fresh-control group ( in vitro oocyte maturation and fertilization, n=9), fresh-transplanted group (ovarian tissue transplantation, in vitro oocyte maturation and fertilization, n=9), frozen-transplanted group (ovarian tissue vitrification-thawed-transplantation, in vitro oocyte maturation and fertilization, n=9). Additionally, frozen-thawed group (ovarian tissue vitrification-thawed, n=6) and ovariectomy group (ovariectomy, n=6) were also set up, to more directly explain the effects of vitrification and freezing on the number of follicles and the changes of endocrine function after ovarian transplantation. We determined the number of follicles by hematoxylin-eosin (HE) staining, neovascularization by CD31 immunohistochemical staining, tissue fibrosis by Masson staining, and serum sex hormone levels by enzyme-linked immunosorbent assay (ELISA) three weeks following ovarian tissue transplantation. In addition, we counted the number of oocytes obtained, oocytes in vitro fertilized, as well as blastocysts. Results:The number of total follicles in both the fresh-transplanted group and the frozen-transplanted group significantly decreased compared with the fresh-control group (all P<0.001) and the frozen-thawed group (all P<0.001). The CD31-positive rate of ovarian tissues in the fresh-transplanted group was significantly higher than that in the fresh-control group ( P=0.044). Although the CD31-positive rate was higher in the frozen-transplanted group than in the fresh-control group, there was no statistical distinction ( P=0.162). The fibrosis area percentage of ovarian tissues in both the fresh-transplanted group and the frozen-transplanted group increased significantly compared with the fresh-control group ( P=0.004; P=0.005). Serum estradiol level in the fresh-transplanted group and the frozen-transplanted group was significantly lower than that in fresh-control group ( P=0.005; P=0.001), significantly higher than that in the ovariectomy group ( P=0.011; P=0.035). Serum follicle-stimulating hormone (FSH) level in the fresh-transplanted group and the frozen-transplanted group was significantly higher than that in the fresh-control group ( P=0.040; P=0.012), significantly lower than that in the ovariectomy group ( P=0.001; P=0.004). In comparison to the fresh-control group, the number of oocytes retrieved in the fresh-transplanted group and the frozen-transplanted group decreased significantly (all P<0.001). Furthermore, the number of oocytes retrieved in the fresh-transplanted group was higher than that in the frozen-transplanted group, and yet there was no statistical difference ( P=0.272). And the number of oocytes in vitro fertilized and blastocysts in the fresh-transplanted group and the frozen-transplanted group were significantly lower than those in the fresh-control group (all P<0.001). The number of total follicles, CD31-positive rate, fibrosis area rate, serum estradiol, and FSH levels, the number of oocytes in vitro fertilized and blastocysts were no substantially distinction between the fresh-transplanted group and the frozen-transplanted group (all P>0.05). Conclusion:?After ovarian tissue vitrification-thawed and transplantation, follicular growth, endocrine function, and fertility were restored in the mouse model, confirming that ovarian tissue vitrification is an effective method for female fertility preservation. Both vitrification and transplantation could cause follicles to be lost and fertility to decrease. And post-transplantation stage is the primary stage of follicle loss during ovarian tissue frozen-thawed-transplantation, and transplantation is the predominant factor affecting the effectiveness of ovarian tissue frozen-thawed-transplantation.
