中国组织工程研究 ›› 2024, Vol. 28 ›› Issue (7): 1050-1056.doi: 10.12307/2024.114

• 干细胞基础实验 basic experiments of stem cells • 上一篇    下一篇

基于第3天胚胎培养液340 nm处吸光度建立优质胚胎预测模型及验证

周  超1,庾广聿1,范嘉琪1,于春梅2,吴  敏1,陈施蓓1   

  1. 1广西壮族自治区南溪山医院,广西壮族自治区桂林市   541000;2常州市妇幼保健院,江苏省常州市   213000
  • 收稿日期:2023-02-06 修回日期:2023-03-23 出版日期:2024-03-08 发布日期:2023-07-17
  • 通讯作者: 于春梅,硕士,副主任医师,常州市妇幼保健院,江苏省常州市 213000
  • 作者简介:周超,男,1992年生,广西壮族自治区灵川县人,汉族,2014年右江民族医学院毕业,主管技师,主要从事辅助生殖胚胎实验室及男科实验室工作,现主要研究方向为辅助生殖临床相关预测模型。
  • 基金资助:


Establishment and validation of embryo high-quality prediction models based on the third-day 340 nm absorbance embryo culture

Zhou Chao1, Yu Guangyu1, Fan Jiaqi1, Yu Chunmei2, Wu Min1, Chen Shibei1   

  1. 1Nanxishan Hospital of Guangxi Zhuang Autonomous Region, Guilin 541000, Guangxi Zhuang Autonomous Region, China; 2Changzhou Maternal and Child Health Hospital, Changzhou 213000, Jiangsu Province, China
  • Received:2023-02-06 Revised:2023-03-23 Online:2024-03-08 Published:2023-07-17
  • Contact: Yu Chunmei, Master, Associate chief physician, Changzhou Maternal and Child Health Hospital, Changzhou 213000, Jiangsu Province, China
  • About author:Zhou Chao, Technologist-in-charge, Nanxishan Hospital of Guangxi Zhuang Autonomous Region, Guilin 541000, Guangxi Zhuang Autonomous Region, China

摘要:


文题释义:

优质胚胎:在辅助生殖技术中,优质胚胎的定义主要基于胚胎形态学评估的结果。胚胎形态学是对胚胎胎龄、卵裂球个数、碎片率等情况进行综合评估的方法。其中第3天优质胚胎定义为卵裂球个数为7-9细胞,Ⅰ,Ⅱ级胚胎评为优质胚胎,Ⅲ,Ⅳ级胚胎为非优质胚胎;优质囊胚定义为≥3期的囊胚且囊胚内细胞团质量和滋养外胚层细胞评分不含C级胚胎3AA、3AB、3BA、3BB的囊胚为优质囊胚,评级至少1个为C级的囊胚为非优质囊胚。
胚胎代谢组学研究:研究胚胎基因与环境交互作用后,通过对相对分子质量低于1 000的小分子代谢产物进行定量分析,以寻找代谢物与生理病理变化的相对关系的研究方式。胚胎代谢组学因无创以及具备代谢产物经生物放大作用后易于检测等优点,现已成为胚胎质量评估的热点。


背景:既往大量研究证实,高浓度代谢产物与胚胎质量及临床结局具有显著相关性,而且胚胎发育沉默理论表明正常发育胚胎体外培养时与外界保持较低的物质交换水平,而当胚胎出现DNA损伤时常因应激修复机制而表现出代谢异常活跃状态。

目的:基于第3天胚胎培养液340 nm处吸光度建立优质胚胎预测模型及验证,为更客观而准确的胚胎质量评估提供依据。
方法:回顾性分析2019年11月至2021年12月于广西壮族自治区南溪山医院接受体外受精-胚胎移植患者269例,其中162例于第3天形成优质胚胎873枚,优质囊胚214枚,将其纳入优质胚胎组;107例于第3天形成非优质胚胎859枚,非优质囊胚214枚,将其纳入非优质胚胎组;采用朗伯-比尔定律筛选出对优质与非优质胚胎具有区分度的特征波长,分析其与优质胚胎的相关性及影响趋势,并建立该波长下吸光度对优质与非优质胚胎的临床预测模型并验证。

结果与结论:①第3天优质与非优质胚胎培养液于340 nm处吸光度差异有显著性意义(P < 0.001),且与第3天优质胚胎的形成呈负相关性(r=-0.486,P < 0.001);优质与非优质囊胚在培养液340 nm处吸光度差异有显著性意义(P < 0.05),且与优质囊胚的形成呈负相关性(r=-0.642,P < 0.001);②受试者工作曲线分析显示,第3天优质与非优质胚胎培养液于340 nm处吸光度的最佳截断值为0.235,曲线下面积为0.799,敏感度为62.9%、特异度为78.0%、准确度为70.5%;优质与非优质囊胚培养液于340 nm处吸光度的最佳截断值为0.175,曲线下面积为0.871,敏感度为74.3%、特异度为89.1%、准确度为82.2%;③限制性立方样条曲线分析显示,当培养液340 nm处吸光度≥0.221时,对第3天非优质胚胎的形成具有显著的正影响趋势,当培养液340 nm处吸光度≥0.160时,对非优质囊胚的形成具有显著的正影响趋势;④临床决策曲线与临床影响曲线显示,培养液340 nm处吸光度对第3天优质胚胎与优质囊胚预测模型分别于阈概率为0.18-0.95与0.16-1.00时具有临床最大净获益,且在该阈概率范围内损失与获益的比值始终小于1,证实该预测模型具有较好的临床应用效能;胚胎移植结果显示,未妊娠患者胚胎培养液于340 nm处吸光度显著高于临床妊娠、生化妊娠、早期流产患者(P < 0.05);⑤结果表明,优质与非优质胚胎培养液于340 nm处吸光度具有显著的差异性与相关性,通过对其建立的优质胚胎预测模型具有一定的临床价值与应用效能,通过联合胚胎形态学评估能一定程度提高胚胎质量评估的客观性与准确性。

https://orcid.org/0000-0001-9843-6731 (周超) 

中国组织工程研究杂志出版内容重点:干细胞;骨髓干细胞;造血干细胞;脂肪干细胞;肿瘤干细胞;胚胎干细胞;脐带脐血干细胞;干细胞诱导;干细胞分化;组织工程

关键词: 胚胎质量评估, 胚胎代谢组学, 胚胎形态学评估, 预测模型, 优质胚胎

Abstract: BACKGROUND: A large number of previous studies have confirmed that a high concentration of metabolites is significantly correlated with embryo quality and clinical outcome, and the theory of silencing embryo development indicates that normally developed embryos maintain a low level of material exchange with the outside world during in vitro culture, while embryos often show abnormal metabolic activity due to stress repair mechanism when DNA damage occurs. 
OBJECTIVE: To establish and verify an embryo quality prediction model based on the third-day 340 nm absorbance embryo cultures to provide the basis for a more objective and accurate embryo quality assessment. 
METHODS: 269 patients at the Nanxishan Hospital of Guangxi Zhuang Autonomous Region for in vitro fertilization and embryo transplantation from November 2019 to December 2021 were retrospectively analyzed. Among them, on day 3, 162 cases who had 873 optimal embryos and 214 high-quality blastocysts were included in the high-quality embryo group. On day 3, 107 cases who had 859 non-optimal embryos and 214 non-high-quality blastocysts were included in the non-high-quality embryo group. Lambert-beer law was used to screen out the characteristic wavelength with distinguishing degree between superior and non-superior embryos, analyze its correlation and influence trend with high-quality embryos, and establish the clinical prediction model and validation of absorbance for high-quality and non-high-quality embryos at this wavelength.
RESULTS AND CONCLUSION: (1) There was a significant difference in absorbance between high-quality and non-high-quality embryos at 340 nm on day 3 (P < 0.001), and a negative correlation was found with the formation of high-quality embryos on day 3 (r=-0.486, P < 0.001). The absorbance of high-quality and non-high-quality blastocyst at 340 nm was significantly different (P < 0.05), and was negatively correlated with the formation of high-quality blastocyst (r=-0.642, P < 0.001). (2) The optimal cut-off value of absorbance at 340 nm between high-quality and non-high-quality embryos on day 3 was 0.235. The area under the curve was 0.799. Sensitivity was 62.9%. Specificity was 78.0%. Accuracy was 70.5%. The optimum cutoff value of high-quality and non-high-quality blastocysts of absorbance at 340 nm was 0.175. The area under the curve was 0.871. Sensitivity was 74.3%. Specificity was 89.1%. Accuracy was 82.2%. (3) Restricted cubic spline curve analysis showed that when the absorbance of the culture medium at 340 nm was greater than 0.221, there was a significant positive trend on the formation of non-high-quality embryos at day 3, and when the absorbance of the culture medium at 340 nm was greater than 0.160, there was a significant positive trend on the formation of non-high-quality blastocysts. (4) The clinical decision curve and clinical influence curve showed that the absorbance of the culture medium at 340 nm had the maximum clinical net benefit for the prediction models of high-quality embryos and high-quality blastocysts on the third day when the valve probability was 0.18-0.95 and 0.16-1.00, respectively, and the ratio of loss to gain within the valve probability range was always less than 1. It is proven that the prediction model has good efficacy in clinical applications. The results of embryo transfer showed that the absorbance of embryo culture medium at 340 nm in non-pregnant patients was significantly higher than that in clinical pregnancy, biochemical pregnancy and early abortion patients (P < 0.05). (5) The high-quality and non-high-quality embryo culture in 340 nm absorbance has a significant difference with correlation. The embryo quality prediction model has a certain clinical value and application effectiveness. The joint embryo morphology evaluation to a certain extent improves the objectivity and accuracy of embryo quality evaluation.

Key words: embryo quality assessment, embryo metabolomics, embryo morphology evaluation, prediction model, high-quality embryo

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