Comparison of the effects of vitrification and programmed freezing protocol on cryopreservation of macaque ovarian tissue
10.3760.cma.j.cn101441-20190318-00104
- VernacularTitle:玻璃化冷冻和程序化冷冻方法保存猕猴卵巢组织的存活效果比较
- Author:
Hua ZHAO
1
;
Yuhui ZHANG
1
;
Lei LI
1
;
Lu WANG
1
;
Jingyu LI
1
;
Linlin LIANG
1
Author Information
1. 河南省人民医院生殖中心，郑州 450000
- Publication Type:Journal Article
- Keywords:
Vitrification freezing;
Programmed freezing;
Ovarian tissue freezing;
Phosphohistone H3
- From:
Chinese Journal of Reproduction and Contraception
2020;40(6):469-475
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the cryopreservation effects of vitrification and programmed freezing protocol on ovarian tissue of macaque.Methods:Fresh macaque ovarian tissues were randomly divided into three groups: control group (fresh ovarian issue), vitrification freezing group and programmed freezing group. HE staining, preantral follicle classification and immunostaining were performed in the three groups. The morphology of follicles, the normal rate of follicles at different stages and the difference in the expression of phosphohistone H3 (PHH3) in three groups of ovarian tissues were compared.Results:1) HE staining showed that there were no significant differences in the normal rate of primary follicles and primordial follicles among the three groups ( P>0.05). The normal rate of single-layer secondary follicles in the programmed freezing group [40.6% (28/69)] was lower than that in vitrification freezing group [53.5% (38/71)] and control group [60.7% (34/56)], and the difference was statistically significant ( P=0.044), but there was no significant difference between vitrification freezing group and control group ( P>0.05). The normal rate of multi-layer secondary follicles in vitrification freezing group [23.4% (11/47) and the programmed freezing group [16.7% (7/42)] was significantly lower than that of control group [57.3%(39/68)] ( P<0.001). The normal rate of multi-layer secondary follicles in vitrification freezing group was higher than that in programmed freezing group, but the difference was not statistically significant ( P>0.05). 2) Comparison of isolated follicles: the normal morphology of primary follicles in the vitrification freezing group and the programmed freezing group was close to control group, the difference was not statistically significant ( P>0.05). The normal rate of secondary follicles in vitrification freezing group [38.2% (34/89)] and programmed freezing group [23.7% (23/97)] was obvious lower than that in control group [56.4% (61/108)]. The difference was statistically significant ( P<0.001). The normal morphology of secondary follicles in vitrification freezing group was significantly higher than that in programmed freezing group ( P<0.001). 3) PHH3 staining: compared with fresh ovarian tissue, the positive expression rate of PHH3 in granulosa cells of vitrification freezing group was close to that of control group ( P>0.05), while the positive expression rate of PHH3 in follicular granulosa cells of programmed freezing group (58.72%±12.31%) was significantly lower than that of control group (67.58%±8.45%, P=0.04) and vitrification freezing group (62.87%±9.94%, P=0.03). Conclusion:Both vitrification and programmed freezing protocol can effectively preserve ovarian tissue, but the morphological damage of follicles and granulosa cells during vitrification and thawing is significantly lower than that of programmed freezing, which is more suitable for the next study of human ovarian tissue cryopreservation.
