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目的探讨在新型玻璃化冷冻法中添加抗凋亡剂是否改善人卵巢组织冷冻保存效果。方法收集2012年10月-2013年8月行妇科手术的患者卵巢标本共10例,使用新型玻璃化冷冻法保存。实验组冷冻保护液中添加抗凋亡剂维生素C,与未添加维生素C组(对照组)比较始基卵泡中卵母细胞和颗粒细胞超微结构、始基卵泡及间质凋亡发生率。结果实验组中卵母细胞和颗粒细胞超微结构正常百分率高于对照组(P<0.05)。在凋亡发生率检测中,实验组和对照组始基卵泡凋亡发生率分别为21.6%(49/227)、38.6%(91/236),差异有统计学意义(P<0.001)。卵巢间质凋亡分析中,实验组和对照组凋亡发生率分别为(21.33±3.41)%、(33.46±3.09)%,差异有统计学意义(P<0.001)。结论在新型玻璃化冷冻法中添加抗凋亡剂能更好地保存始基卵泡及周围间质细胞,改善玻璃化冷冻效果。
Objective To investigate whether the addition of anti-apoptotic agents in the new vitrification method can improve the cryopreservation of human ovarian tissue. Methods A total of 10 ovarian specimens from patients undergoing gynecological surgery from October 2012 to August 2013 were collected and stored in a new vitrification method. The anti-apoptotic agent vitamin C was added into the cryoprotectant solution of experiment group, and the ultrastructure of primordial follicles, primordial follicles and interstitial apoptosis in primordial follicles were compared with those without vitamin C supplementation (control group). Results The normal percentage of oocytes and granulosa cells in the experimental group was higher than that in the control group (P <0.05). In the incidence of apoptosis, the incidence of primordial follicles apoptosis in experimental group and control group were 21.6% (49/227) and 38.6% (91/236), respectively, with significant difference (P <0.001). In ovarian stromal apoptosis analysis, the incidence of apoptosis in experimental group and control group were (21.33 ± 3.41)% and (33.46 ± 3.09)%, respectively, with statistical significance (P <0.001). Conclusion The addition of anti-apoptotic agents in the new vitrification method can better preserve the primordial follicles and the surrounding interstitial cells and improve the vitrification effect.