利用Timelapse技术进行IVF胚胎选择的初步探索docxWord文件下载.docx
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利用Timelapse技术进行IVF胚胎选择的初步探索docxWord文件下载.docx
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4河北农业大学(071000)
【摘要】目的:
通过对早期胚胎发育动态实时监测与影像分析,寻找胚胎发育时期的形态学差异对胚胎发育及后续行为的影响,为胚胎形态学评估及发育潜能预测提供帮助。
方法:
使用Primovision胚胎监测系统,利用延时摄像(timelapse)技术对从2016年3月至2017年2月于东莞市人民医院进行体外受精(invitrofertilization,IVF)治疗中33个周期中收集的308个胚胎进行胚胎实时监测,通过对Day3之前的胚胎发育进行连续检测,系统观察胚胎从原核消失到Day3等不同时间间隔胚胎实时监控图像,结合对卵裂期(2C期)胚胎的评级结果与Day3天胚胎评级结果进行比较;
按照day3胚胎评级结果,分析统计不同等级胚胎从原核期到第一次卵裂及从3细胞期变成4细胞期的时间间隔;
在Day3天进行胚胎评级后,继续对71枚胚胎进行囊胚培养,观察囊胚形成率的差异;
收集胚胎发育时间、事件与形态评分等相关参数,并对囊胚形成率进行对比,寻找上述指标与胚胎发育潜能之间的关系。
结果:
308枚胚胎在Day3获得1级胚胎56枚,2级胚胎109枚,3级胚胎47枚,4级胚胎96枚;
1&
2级胚胎从原核期到卵裂期(Pf-2C)的平均时间为2.28±
0.34h,3级为2.51±
0.42h,4级为2.89±
1.19h;
1&
2级胚胎从3细胞期到4细胞期(3C-4C)的平均时间为0.72±
0.65h,3级为l.14±
1.20h,4级为3.12+3.11h;
结果表明不同胚胎评级间Pf・2C和3C-4C差别均有统计学意义,两两比较采用SNK・Q检验,结果除3级和4级间Pf-2C差别无统计学意义,其他两两间差别均有统计学意义;
对308枚胚胎在第一次卵裂后形成细胞胚胎的形态并进行分类,结果表明,一级2细胞胚胎在Day3形成优质胚胎(1和2级胚胎)的比率为79.4%,二级2细胞胚胎组的为72%,三级2细胞胚胎组为47%,四级2细胞胚胎组的为8.82%,5级二细胞组的为9%,釆用GAMMA法对两有序分类变量进行关联性分析,结果显示细胞胚胎形态与胚胎分级有相关性(G=0.6661,p=0.0392)o71枚胚胎按照Day3胚胎评分分组进行囊胚培养,结果显示1级胚胎囊胚形成率为94.12%,2级为90.91%,3级胚胎为57.14%,4级为5.56%,结果显示,Day3胚胎分级与囊胚形成率差别有统计学意义,两两比较用卡方分割法,并下调检验水准(0.01),1级&
2级(精确p=0.4297)差别无统计学意义;
1级&
3级(精确p=0.0281),差别无统计学意义;
2级&
3级(精确p=0.0361),差别无统计学意义;
4级(精确p=0.0001),差别有统计学意义;
3级&
4级(精确p二0.0037),差别有统计学意义。
姑论:
初步建立了一个便于应用的胚胎选择模型,为IVF实验室的胚胎评估工作提供指导。
【关键词】timelapse;
胚胎选择;
囊胚;
模型
PreliminaryStudyofIVFEmbryoSelectionbyTimelapseTechnology
LiuZhi-gang^ZhangPeng2,QuanYi3,LiangYong-na1,XieSi-sheng\LiXiang-yun4
departmentofReproductiveMedicine,DongguanPeople'
sHospitalofGuangdongProvince,Dongguan523000‘China.
departmentofMarketingandManagement,GuangzhouZunboMedicalDevicesCo.,Ltd,Guangzhou510000,China.
3DepartmentofReproductiveMedicine,JiangmenMaternity&
ChildHealthCareHospital,Jiangmen529000,China.
^ollegeofAnimalScienceandTechnology,AgriculturalUniversityofHebei,Baoding071000,China.
(Abstract]Objective:
Inthisstudy,usingtimelapsetechnologytoanalysisofearlyembryodevelopmentdynamics,theeffectsofmorphologicaldifferencesduringembryonicdevelopmentonembryonicdevelopmentandsubsequentbehaviorsaresearched,tofindtherelationshipbetweentheaboveindicatorsandtheembryodevelopmentpotential.Methods:
thePrimovisionembryomonitoringsystemwasusedtoreal-timemonitor308embryoscollectedin33cycles,andreal-timemonitoringimagesofembryosatdifferenttimeintervalsfromprokaryoticdisappearancetoDay3weresystematicallyobserved.Thescoreofembryosinthephaseofcleavage(2C)werecomparewiththescoreofDay3embryo.Accordingtoday3embryoscoreresults,Thetimeintervalfromprokaryoticstagetofirstcleavagestageandfrom3-cellstageto4-cellstagewereanalyzed.AfterembryogradingonDay3,theblastocystculturewasperformedusing71embryos,toobservetherelationshipbetweenblastocystformationratewithembryogradinglevel.Results:
Within308embryosatDay3,therewere56first-gradeembryos,109second-gradeones,47third-gradeonesand96fourth-gradeones.Themeantimebetweenprokaryoticstagetocleavagestage(pf-2c),1&
2stageembryowas2.28±
0.34h,3stageembryowas2.51±
0.42h,4stageembryowas2.89±
1.19h;
Level1&
2stageembryocellsfrom3to4cells(3cto4c)foranaverageof0.72±
0.65h,level3is1.14±
1.20h,level4is3.12+3.11h,theresultsshowedthatthetimeofpf-2cand3c-4cbetweendifferentembryoratingswerestatisticallysignificant,Thepf-2cdifferencebetweengrade3and4wasnotstatisticallysignificant,whiletheothertwodifferenceswerestatisticallysignificant;
Themorphologyof308embryosformedafterthefirstcleavagewasclassified,theresultshowsthattwocellstagefirst-gradeembryosinDay3formhighqualityembryos(grade1&
2embryo)is79.4%,twocellstagesecondary-gradeembryoswas72%,twocellstagethird-gradeembryoswas47%,twocellstageforth-gradeembryoswas8.82%,twocellstagefifth-gradeembryoswas9%,byusingtwomethodofGAMMAorderlyclassificationvariables,correlationanalysisresultsshowthattheembryonicformhascorrelationwithembryocells(G=0.6661,p=0.6661).Blastocystculturewasperformedon71embryosgroupedaccordingtoDay3embryoscore,andtheresultsshowedthattheblastocystformationrateofstage1embryoswas94.12%,stage2embryoswas90.91%,stage3embryoswas57.14%,andstage4embryoswere5.56%.TheresultsshowedthatthedifferencebetweenDay3embryoclassificationandblastocystformationratewasstatisticallysignificant,Thechi-squaresegmentationmethodwasusedforpairwisecomparison,Grade1comparegrade2(p=0.4297),thedifferencewasnotstatisticallysignificant,Grade1comparegrade3(p=0.0281),thedifferencewasnotstatisticallysignificant.Grade2comparegrade3(p=0.0361),thedifferencewasnotstatis
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