Comparison of short-term transplantation effect of different vitrification solution on human ovarian tissue
10.3760/cma.j.cn101441-20200608-00335
- VernacularTitle:不同玻璃液冻存人卵巢组织的近期异种移植效果的比较
- Author:
Xiaohong LI
1
;
Huiru TANG
;
Yushan LI
;
Yucui ZENG
;
Weixia WEI
;
Bin HUANG
;
Hui DU
;
Ruifang WU
Author Information
1. 北京大学深圳医院，深圳 518036
- Publication Type:Journal Article
- Keywords:
Human ovarian tissue;
Vitrification;
Appoptosis;
Microvessel density
- From:
Chinese Journal of Reproduction and Contraception
2022;42(1):58-64
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the short-term transplantation effect of different vitrification sollutions in order to optimize the cryopreservation protocol on human ovarian tissue.Methods:Ovarian tissues came from 4 patients who were treated surgically due to gynecological diseases in Peking University Shenzhen Hospital from March 2018 to December 2019. The size of 5-10 mm×10 mm×1 mm ovarian tissues were distributed to fresh control group and four experimental groups (G1, G2, G3 and G4). After equilibrium and thawing, the frozen ovarian tissues were grafted into chorioallantoic membrane (CAM). The transplanted ovarian tissues were collected 5 d after transplantation. The morphology of follicles was assessed by HE, the expression of CD105 protein was detected by immunohistochemistry and the microvascular density was recorded. The expression of Bcl-2 and Bax proteins was detected by Western blotting.Results:There was a significantly difference in the percentage of morphologically normal primordial follicles (MNPF) among G1 [63.2% (24/38)], G2 [68.3% (28/41)], G3 [61.9% (26/42)], G4 [69.6% (32/46)] and fresh control groups [82.8% (82/99), P=0.044]. Compared with fresh control group, the percentages of MNPF were statistically lower in the treatment groups (all P<0.05). The CD105-microvascular density (MVD) in fresh control group, G1, G2, G3 and G4 groups were 6.51±1.30/mm 2, 11.10±1.62/mm 2, 13.04±1.84/mm 2, 9.11±1.09/mm 2, 11.28±1.62/mm 2, respectively. The CD105-MVD in four treatment groups after transplantation was significantly higher than that of fresh control group ( P<0.001, P<0.001, P=0.022, P<0.001), and the CD105-MVD of G1, G2 and G4 groups was higher than that of G3 group ( P=0.024, P<0.001, P=0.034). The ratio of Bcl-2/Bax in G1, G2, G3, G4 groups and fresh control group were 0.71±0.37, 0.84±0.29, 0.45±0.18, 0.84±0.29 and 0.44±0.21, respectively. The ratio of Bcl-2/Bax of G2 group was statistically higher than that of fresh control group and G3 group ( P=0.025, P=0.038). Conclusion:After 5 d of transplantation into CAM, the percentage of normal primordial follicles is lower. Acorrding to the higher vascularization and anti-apoptotic ability, G2 and G4 are the optimal vitrification protocol for human ovarian tissue cryopreservation.
