用11色探针检测植入前胚胎非整倍体的实验研究

Experimental study of embryo's aneuploidy using 11 different probes

目的建立用11种不同的染色体探针进行三轮FISH杂交检测单卵裂球的技术,同时探讨胚胎中的嵌合体现象及1PN胚胎的整倍性。方法收集IVF/ICSI治疗周期中,受精后第3天的废弃胚胎31枚,包括15枚2PN胚胎和16枚1PN胚胎用于研究。蛋白酶消化透明带,每个胚胎中吸取2、3个卵裂球,用0.2%Tween-20/0.01N HCl+甲醇/冰醋酸(3∶1)法固定,用11种染色体探针进行FISH杂交分析。结果共获72个卵裂球,成功固定核69个,成功率为95.8%;FISH杂交后,66个核信号完整,杂交成功率为95.6%;9个胚胎为嵌合型,嵌合体率为29.0%;16个1PN胚胎中6个正常,正常率为37.5%。结论建立了较稳定的应用11色探针进行FISH杂交诊断单卵裂球技术;第3天胚胎中嵌合体率较高,会增加PGS的误诊率;部分1PN胚胎染色体正常,在无2PN胚胎移植的情况下,1PN胚胎也可作为选择。

[ Objective ] To establish the PGS technology using 11 different FISH probe to test blastomere, and to investigate the mosaic rate in embryos and aneuploidy of 1Pnembryos. [Method] 31 D3 embryos discarded in IVF/ ICSI cycles, including 15 2PN embryos and 16 1PN embryos were collected. Digested zona pellucida and got 2~3 blastomere from every embryo, fixed blastomere with 0.2% Tween-20/O.O1N HCl＋methanol/acetic acid （3：1）, and then analyzed using 11 different probes. [ Result ] 72 blastomeres were obtained. 69 nuclears were fixed and the rate was 95.8%. After FISH, 66 nuclear had complete signals, the rate was 95.6%. 9 embryos were mosaic, the rate was 29.0%. 6 out of 16 1PN embryos were normal, the rate was 37.5%. [Conclusion] We established the stable FISH technology using 11 different probes to diagnose blastomeres. The mosaic rate of D3 embryos was high and it would improve misdiagnosis rate of PGS. A part of 1PN embryos had normal chromosome, so when 2PN embryos can not be obtained, 1PN embryos could be a choice for transplant.