未成熟卵母细胞体外成熟不同培养结局的多因素分析

Multivariate analysis of factors for outcomes of in vitro maturation culture of human immature oocytes

目的应用多因素分析探讨人未成熟卵母细胞进行体外成熟(IVM)培养后不同培养结局(卵母细胞成熟和D3可利用胚胎)的影响因素。方法收集2020年6月至2023年6月于中山市人民医院生殖中心进行常规助孕治疗、有未成熟卵母细胞剩余患者的临床资料。按照纳排标准共纳入45例患者的207枚未成熟卵母细胞,使用商品化IVM培养液和自配IVM培养液进行IVM培养。培养24 h和48 h后,观察卵母细胞成熟情况。当卵母细胞成熟后,行常规卵胞浆内单精子注射(ICSI)、胚胎培养和胚胎观察,记录D3可用胚胎数。并采用Logistic回归分析不同培养液、女性年龄、不孕因素、体质量指数(BMI)等因素对卵母细胞成熟率和D3可用胚胎率的影响。结果纳入研究的207枚未成熟卵母细胞进行IVM培养后,获得成熟卵母细胞159个,成熟率为76.81%;获得D3可用胚胎73个,D3可用胚胎率为35.27%。不同IVM培养液、女方年龄、促排卵方案、BMI、基础FSH(bFSH)、基础LH(bLH)、FSH/LH比值、抗苗勒管激素(AMH)、不孕年限、不孕类型、不孕因素及未成熟卵状态的卵母细胞成熟率无显著性差异(P>0.05)。使用商品化IVM培养液的D3可用胚胎率为42.86%,显著高于使用自配IVM培养液的27.45%(P<0.05);不同女方年龄、促排卵方案、BMI等因素的D3可用胚胎率无显著性差异(P>0.05)。多因素Logistic回归分析显示,IVM培养液、女方年龄等都不是卵母细胞成熟的独立影响因素;而IVM培养液和女方年龄是D3可用胚胎率的独立影响因素(P<0.05)。结论经HCG刺激的未成熟卵母细胞进行IVM培养时,使用商品化IVM培养液比使用自制培养液可能更有利于后续D3可利用胚胎的形成;且随着女方年龄增加,IVM成熟卵母细胞来源的D3可用胚胎率下降。

Objective:To explore factors for outcomes of in vitro maturation(IVM)culture of human immature oocytes(oocyte mature and day-3 available embryos)with multivariate factors analysis.Methods:The clinical data of patients with immature oocytes who underwent IVF/ICSI treatment at the Assisted Reproductive Center of Zhongshan City People’s Hospital from June 2020 to June 2023 were collected.In accordance with the inclusion and exclusion criteria,a total of 207 immature oocytes from 45 patients were included in this study.IVM culture was conducted using both commercial IVM culture medium and self-made IVM culture medium.After 24 and 48 hours of culture,the status of the oocytes was assessed.Once oocytes maturing,ICSI,embryo culture and observation of the embryos were performed.The quantity of day-3 available embryos was documented.Then,logistic regression analysis was used to evaluate the influencing factors for the rates of oocyte maturation and day-3 available embryos,such as culture media,female age,infertility factors and body mass index(BMI)and so on.Results:A total of 207 immature oocytes were cultured for IVM in this study,resulting in 159 mature oocytes,with the mature rate of 76.81%,and 73 day-3 available embryos,with the day-3 available embryo formation rate of 35.27%.There was no significant difference in oocyte maturation rate among various IVM culture media,female age,ovulation induction protocols,BMI,basal FSH levels,basal LH levels,FSH/LH ratio,anti-Müllerian hormone(AMH)levels,years of infertility,types of infertility,factors for infertility,and status of immature oocytes(P>0.05).The rate of day-3 available embryos using commercial IVM culture medium was 42.86%,which was significantly higher when compared to self-made IVM culture medium(27.45%,P<0.05).There was no significant difference in the rate of day-3 available embryos among various female age,ovulation induction protocols,BMI and so on(P>0.05).A multivariate logistic regression analysis revealed that factors,such as IVM culture medium,female age and so on,were not independent factors for oocyte maturation.Nonetheless,IVM culture medium and female age were found to be independent factors influencing the rate of day-3 available embryos(P<0.05).Conclusions:When immature oocytes are triggered with HCG and undergo IVM,the use of commercial IVM medium may be more favorable for the formation of day-3 available embryos compared to self-made medium.Moreover,as the female age increases,the rate of day-3 available embryos derived from IVM mature oocytes,shows a decrease.