小鼠卵母细胞慢速冷冻的初步研究——附人妊娠分娩1例报道

Study of Slow Cooling Cryopreservation Protocols of Mouse Oocyte——One Birth of a Healthy Human Baby

目的:探讨不同浓度蔗糖联合丙二醇(PROH)冷冻保护剂对小鼠卵母细胞冻融的影响。方法:①观察不同蔗糖浓度(0.1mol/L、0.2mol/L、0.3mol/L)联合应用1.5mol/L的PROH对小鼠卵母细胞复苏率的影响。②对1例采卵日未能取得精子的不孕症患者,应用0.3mol/L蔗糖联合1.5mol/LPROH将其卵母细胞用慢速冷冻方法冷冻保存,随后解冻并移植。结果:①随着蔗糖浓度的提高,卵母细胞复苏率也随之升高,0.3mol/L蔗糖组复苏率达74%。②1例患者卵母细胞复苏后行卵胞浆内ICSI-ET,获临床妊娠,妊娠中期羊水染色体核型分析正常,妊娠35周因妊高征剖宫产1健康男婴。结论:对卵母细胞慢速冷冻中,适当提高蔗糖浓度可获得较满意的复苏率、受精率及卵裂率,胚胎移植后能够出生健康新生儿。

Objective： To investigate the effects of different sucrose concentrations in a slow-freezing method on the mouse oocyte survival. Methods： （1）The effects of different sucrose concentrations （0.1 mol/L, 0.2 mol/L, 0.3 tool/L） combined with PROH on the mouse oocyte survival was observed. （2） To one patient undergoing ICSI cycle and no sperm was achieved, 1.5 mol/L PROH＋0.3 mol/L sucrose was used to preserve all of her mature oocytes by slow-freezing method. Results： （1） The survival rate of mouse oocytes increased with the sucrose concentration increased, 0.3 mol/L sucrose in the cryoprotectant resulted in survival rate of 74%. （2） The human mature oocytes were thawed and ICSI was performed, 2 embryos were transfered, clinical pregnancy was achieved, amniocentesis was performed at 19 weeks of gestation, fatal chromosomes showed normal, a healthy, normal male infant was born by cesarean section at 35 weeks＇ gestation because of the pregnancy-inducedhypertention. Conclusion： The results suggest that sucrose concentration of 0.3 mol/L in the cryoprotectant solution is efficient in freezing oocytes with slow-freezing method. Slow cooling method for oocyte cryopreservation offers high rates of survival, fertilization, and normal pregnancy, providing a new alternative for the treatment of infertility.