| Objective:By analyzing the differences in morphology and morphokinetic between euploidy embryos and aneuploidy embryos of IVF-ET cycle,combined with the report of preimplantation genetic testing,the influence of individual chromosome aneuploidy on preimplantation stage embryos was explored.At the same time,the prediction model of embryonic euploidy based on embryo morphology and morphokinetic was established to provide a basis for clinical effective selection of embryos and to improve the reproductive safety.Method:The morphological and morphokinetic parameters of the embryo that was cultured in the Embryo Scope plus incubator in the Reproductive Center of the First Affiliated Hospital of Hainan Medical University from June 2019 to September 2022 and trophoblast cells were taken for preimplantation genetic testing were retrospectively analyzed.Statistical methods such as Rank sum test,Chi-square test,Binary Logistic regression and Linear regression were used to analyze the data,to find out the parameters that can reflect the differences between euploidy embryos and aneuploidy embryos,so as to explore the influence of individual chromosome aneuploidy on preimplantation stage embryos.The predictive model of embryonic euploidy was established by using these parameters,ROC curve was drawn,and cut-off values of parameters were found by using Youden index.Results:1.Basic information:A total of 1888 embryos were collected,of which euploid embryos accounted for 38.30%and aneuploid embryos accounted for 61.70%.2.Morphological parameters:(1)Groups were grouped according to D1 prokaryotic score Z1 to Z4,and the Chi-square test of aneuploidy rate among groups showed P>0.05.(2)The degree of cell fragmentation of D2 and D3 was divided into three groups according to 0%～5%,6%～20%and 21%～50%,and the Chi-square test of aneuploidy rate among groups showed P>0.05.(3)The cell homogeneity of D2 and D3 was divided into groups A,B and C,and the Chi-square test of aneuploidy rate among D2 group showed all P>0.05.When the female age was ≥35y,the Chi-square test of the aneuploidy rate among D3 group showed P>0.05;But when the female age was<35y,the aneuploidy rate of group A,group B and group C was 32.79%,56.74%and 54.05%,respectively,group A and group B showed P<0.05,however,P>0.05 was found between group A and group C,and between group B and group C.(4)The blastocysts were divided into three groups according to Gardner ratings of inner cell mass and trophoblast cells:excellent group=AA,good group=AB,BA,BB,fair group=AC,BC,CA,CB.When the female age was<35y,the aneuploidy rates of the three groups were 46.76%,54.90%,68.21%,respectively.P>0.05 between excellent group and good group,P<0.05 between excellent group and good,excellent group and fair group.When the female age was≥35y,the aneuploidy rates of the three groups were 44.22%,76.79%and 84.25%,respectively,P<0.05 between excellent group and good group,excellent group and fair group,and P>0.05 beyween the good group and fair group.3.Morphokinetic parameters:(1)The rank-sum test of morphokinetic parameters of single chromosome aneuploid embryos and euploid embryos,showed that P>0.05 between euploid embryos and embryos with single chromosome aneuploidy such as chromosome 3,chromosome 7,chromosome 8,chromosome 10,chromosome 11,chromosome 12,chromosome 13,chromosome 15,chromosome 18,chromosome 20 and chromosome 21.(2)Before confounding factors were excluded,the rank sum test results of morphokinetic parameters of euploid embryos and aneuploid embryos suggested that P<0.05 for t3,t4,t5,t6,t7,t8,CC3,tSB,tB,tB-t8,tB-tSB,KIDScore,iDAScore,etc.When confounding factors were controlled by multivariate regression analysis,only P<0.05 for t5,CC3,tB,KIDScore,iDAScore.4.Prediction model of embryonic euploidy:The AUCs of t5,CC3,tB,female age,Gardner score,KIDScore and iDAScore were 0.541,0.560,0.593,0.575,0.638,0.624 and 0.648,respectively.When combined with the above parameters,The AUC was 0.686(95%CI 0.660-0.711).5.The euploidy rates of groups according to cut-off values:(1)The euploidy rates of t5≤48.74h and t5>48.74h were 47.22%and 39.55%,respectively,P<0.05.(2)The euploidy rates of CC3≤20.27h and CC3>20.27h were 47.50%and 35.4%,respectively,P<0.05.(3)The euploidy rates of tB≤111.74h and tB>111.64h were 48.48%and 32.43%,respectively,P<0.05.(4)The euploidy rates of KIDScore≥5.65 and KIDScore<5.65 were 52.15%and 31.79%,respectively,P<0.05.(5)The euploidy rates of iDAScore≥8.45 and iDAScore<8.45 were 53.46%and 29.22%,respectively,P<0.05.Conclusion:1.In embryos that has become blastocysts,aneuploidy of embryonic genome had no effect on morphology of D1 and D2 embryos,had fair effect on D3 embryos.But had significant effect on blastocysts,the worse the Gardner score,the higher the aneuploidy rate.2.The influence of aneuploidy on the morphokinetic of the embryo starts from t5,and the development of euploid embryos is faster than that of aneuploid embryos.3.No evidence show that single chromosome aneuploidy such as chromosome 3,chromosome 7,chromosome 8,chromosome 10,chromosome 11,chromosome 12,chromosome 13,chromosome 15,chromosome 18,chromosome 20 and chromosome 21 have influence on morphokinetic of embryos from fertilization to preimplantation stage.4.t5,CC3,tB,female age,Gardner score,KIDScore and iDAScore score were included in the model to predict embryonic euploidy,and the AUC of ROC curve was 0.686(95%CI 0.660-0.711).5.When the embryos’ t5>48.74h,CC3>20.27h,tB>111.64h,KIDScore<5.65,iDAScore score<8.45,the embryonic euploidy rate is lower,if the patient has a fair number of embryos,the selection of these embryos for genetic testing or transplantation should be avoided. |
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