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目的:建立同时将13、18、21、X、Y五色探针用于分析单个卵裂球的方法。方法:收集体外受精-胚胎移植治疗周期中受精第3天,不适宜移植和冷冻的正常受精胚胎11个、双精受精胚胎1个用于研究。用Tyrode′s酸溶解这些胚胎的透明带后,用0.1%Tween-20/0.01NHCl将卵裂球固定,用Vysis公司的MultiVysion PGT FISH探针(含有13、18、21、X、Y五种染色体探针)进行分析。结果:12个胚胎共有46个卵裂球,有核的卵裂球43个,其中有3个卵裂球各有2个核,共有46个细胞核。成功固定45个核,固定成功率为97.8%。43个核有杂交信号,杂交率为95.6%。11个单精受精胚胎共有39个核有信号,都为二倍体胚胎,其中正常胚胎2个。在9个异常胚胎中,嵌合型胚胎6个,异常非嵌合型胚胎2个,无规律分离型1个。结论:种植前胚胎中,染色体异常胚胎广泛存在,非整倍体胚胎占染色体异常胚胎的绝大多数,嵌合型是非整倍体的主要形式,形成非整倍体的主要原因可能是减数分裂过程中染色体丢失。
OBJECTIVE: To establish a method for simultaneous analysis of single blastomeres with 13, 18, 21, X, Y five-color probes. Methods: Eleven fertilized embryos were collected on the third day after in vitro fertilization-embryo transfer (IVF-ET) cycle, which were unsuitable for transplantation and freezing. One double-fertilized embryo was used for the study. After the zona pellucida of these embryos was lysed with Tyrode’s acid, the blastomeres were fixed with 0.1% Tween-20 / 0.01NHCl and analyzed with Vysis’s MultiVysion PGT FISH probe (containing 13, 18, 21, X, Chromosome probe) for analysis. RESULTS: There were 46 blastomeres and 43 nuclear blastomeres in 12 embryos, of which 3 had 2 nuclei and 46 nuclei in total. Successfully fixed 45 nuclei, a fixed success rate of 97.8%. 43 nuclear hybridization signals, the rate of 95.6% hybridization. A total of 39 nuclear signals were detected in 11 single fertilized embryos, all of which were diploid embryos, including 2 normal embryos. Among 9 abnormal embryos, 6 were chimeric embryos, 2 were abnormal non-chimeric embryos, and 1 was irregularly segregated. Conclusion: Among the preimplantation embryos, there are many abnormal embryos, aneuploid embryos make up the vast majority of aneuploid embryos, and chimeric ones are the main forms of aneuploidy. The main reason for aneuploidy may be subtraction Chromosomes are lost during division.