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目的:探讨着床前遗传学诊断(PGD)后囊胚冻融的特点及临床应用。方法:①实验研究:对41枚PGD诊断为遗传学异常的废弃囊胚进行冻融研究,其中27枚囊胚进行程序化冻融,14枚囊胚进行玻璃化冻融。分别观察囊胚冷冻前及解冻后的形态学特点,比较2种冻融方法的囊胚解冻复苏率。②临床应用:对14对夫妇在PGD新鲜周期移植后仍有剩余的遗传诊断正常的40枚囊胚给予冷冻保存。其中,6对夫妇的12枚胚胎进行了解冻移植。结果:①实验研究:27枚进行程序化冻融的囊胚解冻后复苏22枚,复苏率为81.48%(22/27);14枚进行玻璃化冻融的囊胚解冻后复苏11枚,复苏率为78.57%(11/14),2种冻融方法的囊胚解冻复苏率无统计学差异(P>0.05)。无论何种冻融方法,解冻后复苏存活的囊胚在体外培养30 min均可以观察到复苏现象,体外培养4 h囊腔均扩张。②临床应用:14对有剩余囊胚冻存的夫妇中,6对夫妇新鲜周期即获得临床持续妊娠,另外还有1对夫妇早期自然流产,1对夫妇宫外孕。其余新鲜周期未孕的6对夫妇进行了囊胚解冻移植,其中3对夫妇解冻周期获得了临床妊娠。结论:经PGD诊断后的囊胚,无论是程序化冻融还是玻璃化冻融,均可以获得满意的解冻复苏率。PGD诊断后囊胚冻融技术的成熟与发展可以提高PGD夫妇的累积妊娠率。
Objective: To investigate the characteristics and clinical application of post-bed genetic diagnosis (PGD) post-blastocyst freezing and thawing. Methods: ①Experimental study: Forty-one aborted blastocysts with PGD diagnosed as genetic abnormalities were subjected to freeze-thaw cycles. 27 blastocysts were programmed freeze-thaw and 14 blastocysts were vitrified and thawed. The morphological characteristics of the blastocysts before and after thawing were observed, and the blastocyst thawing and thawing rates of the two kinds of freezing and thawing methods were compared. ② Clinical application: Forty fourty couples of blastocysts with the remaining genetically diagnosed normal after PGD fresh-cycle transplantation were cryopreserved. Among them, 12 couples of 6 embryos were thawed. Results: ①Experimental study: Twenty-seven blastocysts thawed by programmed thawing were resuscitated after resuscitation, the recovery rate was 81.48% (22/27). In the 14 frozen-thawed blastocysts, 11 thawed blastocysts recovered and 11 recovered The rate of blastocyst thawing was 78.57% (11/14). There was no significant difference in blastocyst thaw resurgence between the two freezing and thawing methods (P> 0.05). No matter what kind of freeze-thaw method, the thawed blastocysts survived thawing could be observed after 30 min of in vitro resuscitation, and all the cysts were expanded 4 h after in vitro culture. ② Clinical application: Among 14 couples with remaining blastocysts, 6 couples got clinical continuous pregnancies in their fresh cycles, while 1 couples had early spontaneous abortion and 1 couple had ectopic pregnancy. The remaining 6 couples who had no fresh cycle had blastocyst thawed, of whom 3 had clinical pregnancy during the thaw cycle. Conclusion: The PGD-diagnosed blastocysts, whether procedural freeze-thaw or vitrification freeze-thaw, can achieve satisfactory thaw resuscitation rates. PGD diagnosis of blastocyst after the maturation and development of technology can improve the PGD couple’s cumulative pregnancy rate.