人卵巢组织慢速冷冻保护液中添加抗凋亡剂研究

Investigation on the Effect of Adding Antioxidant Supplementation into the Cryoprotectant Solution on Slow Freezing of Human Ovarian Tissue

目的探讨在人卵巢组织慢速冷冻保护液中添加抗凋亡剂对人卵巢组织的冷冻保存效果。方法收集2012年3月-2013年4月行妇科手术的患者卵巢组织共9例,使用慢速冷冻法保存。实验组冷冻保护液中添加抗凋亡剂维生素C,并与未添加抗凋亡剂组(对照组)比较始基卵泡形态、始基卵泡及间质细胞凋亡发生率。结果实验组中始基卵泡形态正常百分率高于对照组(P<0.05);在凋亡发生率检测中,实验组与对照组相比较,始基卵泡和间质细胞凋亡发生率降低(P<0.05)。结论慢速冷冻法中,冷冻保护液中添加抗凋亡剂能更好地保存始基卵泡及周围间质细胞,在深低温保存人卵巢组织中具有推广价值。

Objective To determine if the cryoprotectant solution supplementation with Vitamin C can improve the protective effect of human ovarian tissue cryopreservation by anti-apoptosis. Methods Human ovarian cortical tissues were collected from nine patients treated between March 2012 and April 2013. The cortical tissue pieces obtained from each patient were divided into two groups： control（conventional slow freezing） and trial group（slow freezing supplementation with Vitamin C）. The preservation effects in the two groups were compared by histology using light microscope and apoptosis assessed by TUNEL assay. Results The proportion of morphologically normal primordial follicles in the trial group was higher than that in the control group（P〈0.05）. The proportion of apoptotic primordial follicles and stromal cells in the trial group was lower than that in the control group（P〈0.05）. Conclusions Cryoprotectant solutions supplementation with Vitamin C can improve the preservation of primordial follicles and stromal cells. It might be a method worth to try in order to improve the protective effect of human ovarian tissue cryopreservation by inhibiting of apoptosis.