冻融人卵子胞浆内单精子注射临床妊娠成功

Successful pregnancy derived from slowly freezing donated human oocytes by intracytoplasmic sperm injection

目的 提供一种适合于临床应用的缓慢低温冷冻和快速解冻复苏的贮存人卵母细胞的方法。 方法 接受试管婴儿 (IVF)或卵子胞浆内单精子注射 (ICSI)治疗的 6例患者剩余的卵母细胞共 33个 ,通过程序化缓慢低温冷冻方法贮存于含有蔗糖的 1,2丙二醇中。 6例因不同原因需要受卵者的子宫内膜用激素替代治疗 (HRT)法调整或采用自然周期 ,当受卵者子宫内膜处于合适时机时 ,复苏冻存的卵母细胞 6次共 33个 ,共 2 8个成熟卵子通过ICSI受精 ,记录卵母细胞复苏率、复苏卵ICSI后的受精率 ,及随后胚胎发育情况 ,选择质量好的胚胎进行移植 ,胚胎移植后 2周检查受孕情况 ,孕 4周B超检查。 结果 当用高浓度的蔗糖冻存液保存时 ,慢冻快融法得到了很高的存活率。复苏后的卵子存活率 97% (32 / 33) ,复苏卵经ICSI后的受精率 82 % (2 3/ 2 8) ,受精胚胎分裂率 91% (2 1/ 2 3) ,6个胚胎移植周期共移植胚胎 2 1个。 4例获临床妊娠 ,目前 1例孕 17周 ,1例孕 12周 ,1例孕 7周 ,均为单胎妊娠 ,状态良好 ;另外 1例妊娠 9周后自然流产。 结论 人卵母细胞慢冻快融低温贮藏方法能够提高人卵母细胞冻存后的存活率。该法操作简便、费用低、复苏率高 ,较好地解决了现有人卵母细胞的技术冻存效果不理想、复苏率低的问题 ,为进一步人卵?

Objective To set up a novel clinical method for storage of oocytes by slowly freezing and successful pregnancy by intracytoplasmic sperm injection (ICSI). Methods The frozen oocytes (n=33) in metaphase Ⅱ (28),which were normal in morphology,were denuded of their cumulus-corona complex.The protective freezing solution contained 1,2-propanediol (1.5 mol/L) and sucrose (0.35 mol/L) and the process was slow for freezing and rapid for thawing.The recipients of these oocytes received hormone replacement therapy (HRT). Results A high survival rate of oocytes (32/33) was obtained when the sucrose concentration was 0.35 mol/L.ICSI induced a higher fertility rate (23/28),a good embryo cleavage rate (21/23) and a satisfactory embryo morphology for thawed oocytes.Embryo transfers were performed in 6 cycles/6 cases. Four cases got clinical pregnancy by demonstration of four gestation sacs on B ultrasonic examination 7 weeks after ICSI,1 fetal was first trimester spontaneous abortion,another 3 survival fetal were ongoing pregnancy and the first case gestation week is 17 weeks. Conclusions This study is the first successful application of human oocyte cryopreservation in China and this method can increase the survival rate of freezing oocytes.It’s easy to carry out,low in cost and high in the recovery rate of oocytes after thawing.