不同玻璃液冻存人卵巢组织的近期异种移植效果的比较

目的:通过研究不同的玻璃液对人卵巢组织冻存后的近期异种移植效果，以筛选出较优的玻璃化冷冻方案。方法:收集2018年3月至2019年12月期间在北京大学深圳医院因妇科疾病行手术治疗的4例患者卵巢组织，将卵巢组织分割成5~10 mm×10 mm×1 mm的小块，分为新鲜对照组和4个处理组，4个处理组分别放入G1、G2、G3...

Comparison of short-term transplantation effect of different vitrification solution on human ovarian tissue

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/mm²,11.10±1.62/mm²,13.04±1.84/mm²,9.11±1.09/mm²,11.28±1.62/mm²,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.