未见原核受精卵衍生胚胎患者的胚胎发育和临床结局

范咏琪; 杨丹丹; 张琦琦; 严博; 章志国

doi:10.16766/j.cnki.issn.1674-4152.003745

未见原核受精卵衍生胚胎患者的胚胎发育和临床结局

doi: 10.16766/j.cnki.issn.1674-4152.003745

1.
安徽医科大学第一附属医院妇产科生殖医学中心，安徽合肥 230032
2.
安徽医科大学第二临床学院，安徽合肥 230032

基金项目:

国家重点研发计划项目 2022YFC2703000

国家自然科学基金项目 82071724

详细信息

通讯作者:
章志国, E-mail: zzg_100@163.com

中图分类号: R321.1 R711.6
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- 文章访问数: 14
- HTML全文浏览量: 8
- PDF下载量: 1
- 被引次数: 0
出版历程
- 收稿日期: 2024-03-17
- 网络出版日期: 2024-12-31

Embryonic development and clinical outcomes in patients with 0PN zygote derived embryos

1.
Reproductive Medicine Center, Department of Obstetrics and Gynecology, the First Affiliated Hospital of Anhui Medical University, Hefei, Anhui 230032, China

摘要

摘要: 目的未见原核(0PN) 受精卵的受精及胚胎发育异常的病因尚不清楚，其移植的适宜性仍存在争议，本研究旨在探究0PN受精卵是否具有应用价值。方法分析2019年9月—2022年5月行体外受精(IVF)/卵胞浆内单精子注射(ICSI)的9 167个周期。(1)比较6 915个周期[每个周期中存在≥1个0PN受精卵，以下简称两原核(2PN)+0PN组]与2 252个周期(每个周期均由2PN受精卵发育而来，以下简称2PN组)的胚胎发育与临床结局。(2)根据0PN衍生胚胎的比例将IVF周期中患者进一步分类为：L组(0~30%，2 579例)、M组(31%~60%，782例)和H组(61%~100%，221例)，比较3组的胚胎发育与临床结局。(3)单独比较72个由0PN受精卵发育周期(0PN100%组)与100个由2PN发育周期(2PN100%组)的胚胎发育与临床结局。结果 (1) 2PN组的囊胚形成率显著高于2PN+0PN组(62.44% vs. 55.01%，P＜0.001)，2PN组的累计活产率(CLR)显著低于2PN+0PN组(54.97% vs. 64.99%，P＜0.001)。2PN+0PN组与2PN组新生儿体重、Apgar评分比较差异均无统计学意义。(2)L、M和H组的获卵数、受精率、累计临床妊娠率(CCR)、CLR呈递减趋势。M组的卵裂率显著高于其余2组；优质胚胎率、囊胚形成率、优质囊胚率显著高于L组。(3)0PN衍生胚胎(0PN100%组)的活产率显著低于2PN衍生胚胎(2PN100%组，30.56% vs. 60.00%，P＜0.001)，但2组囊胚移植后的临床结局差异均无统计学意义(P＞0.05)。结论 0PN衍生胚胎临床妊娠后，并未观察到流产可能性明显增加或对新生儿健康产生明显负面影响。因此，当无法获得正常囊胚时，0PN衍生胚胎的移植应被视为可行的选择。
- 未见原核受精卵 /
- 两原核受精卵 /
- 胚胎发育
Abstract: Objective The cause of abnormal fertilization and embryo development in non-pronucleus (0PN) zygotes remains unclear, and the viability of transferring 0PN zygotes is still controversial. The study aims to explore the clinical utility of 0PN zygote-derived embryos. Methods A total of 9 167 IVF/ICSI cycles conducted between September 2019 and May 2022 were analyzed. (1) Embryo development and clinical outcomes were compared between 6 915 cycles with at least one OPN fertilization (2PN/0PN) and 2 252 cycles with all embryos from 2PN fertilization (2PN group). (2) Patients were further categorized based on the proportion of 0PN-derived embryos in IVF cycles: L group (0-30%, n=2 579), M group (31%-60%, n=782), and H group (61%-100%, n=221), with embryo development and clinical outcomes compared across groups. (3) Embryo development and clinical outcomes were also compared with complete 0PN (0PN 100% group) and complete 2PN (2PN 100% group). Results (1) The blastocyst formation rate of 2PN group was significantly higher than that of the 2PN/0PN group (62.44% vs. 55.01%, P < 0.001), whereas the cumulative live birth rate (CLR) of 2PN group was significantly lower than the 2PN/0PN group (54.97% vs. 64.99%, P < 0.001). There was no significant difference in neonatal weight or Apgar score between the two groups. (2) The number of oocytes retrieved, fertilization rate, cumulative clinical pregnancy rate (CCR), and CLR decreased progressively across L, M, and H. The cleavage rate of group M was significantly higher than that in the other two groups. H group had significantly higher rates of high-quality embryos, blastocyst formation, and top-grade cysts compared to the group L. (3) The live birth rate for 0PN-derived embryos (0PN 100% group) was significantly lower than for 2PN-derived embryos (2PN 100% group, 30.56% vs. 60.00%, P < 0.001), though there was no significant difference in clinical outcomes after blastocyst transfer between the two groups (P>0.05). Conclusion Clinical pregnancy outcomes after transferring 0PN-derived embryos did not show increased risk of miscarriage or adverse neonatal. Therefore, transferring 0PN-derived embryos may be a feasible option when normal blastocysts are unavailable.
- Zygotes with no pronuclei /
- Zygotes with two pronuclei /
- Development of embryos

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表 1 2PN+0PN组和2PN组接受ART治疗患者的基本临床资料比较

Table 1. Comparison of basic clinical data of patients treated with ART between 2PN+0PN group and 2PN group

项目	2PN+0PN组	2PN组	统计量	P值
总周期数(个)	6 915	2 252
BMI(x±s)	22.33±8.50	22.15±3.23	1.233^a	0.327
FSH[M(P₂₅, P₇₅), IU/L]	7.24 (6.00, 8.73)	7.76 (6.46, 9.66)	-10.080^b	＜0.001
LH[M(P₂₅, P₇₅), IU/L]	4.670(3.310，6.590)	4.390(3.173，5.960)	5.391^b	＜0.001
Gn时间(x±s，d)	10.64±2.07	10.25±2.47	7.037^a	＜0.001
Gn剂量[M(P₂₅, P₇₅), IU/L]	2 075.00 (1 650.00，2 700.00)	2 250.00(1 737.500，2 875.00)	-4.989^b	＜0.001
女方年龄[M(P₂₅, P₇₅), 岁]	31 (28, 34)	32 (29, 37)	-9.243^b	＜0.001
原发不孕比例(%)	52.10(3 603/6 915)	49.33(1 111/2 252)	5.219^c	0.022
PCOS比例(%)	17.05(1 179/6 915)	7.37(166/2 252)	127.100^c	＜0.001
子宫内膜异位症比例(%)	2.86(198/6 915)	4.09(92/2 252)	8.280^c	0.004
注：^a为t值，^b为Z值, ^c为χ²值。

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表 2 0PN衍生胚胎发生相关因素的多因素logistic回归分析

Table 2. Multivariate Logistic regression analysis of factors associated with 0PN embryogenesis

变量	B	SE	Waldχ²	P值	OR(95% CI)
FSH	-0.149	0.010	237.428	＜0.001	0.861(0.845~0.878)
LH	0.036	0.009	16.765	＜0.001	1.036(1.019~1.054)
Gn时间	0.068	0.016	18.931	＜0.001	1.070(1.038~1.103)
Gn剂量	0.000	0.000	16.881	＜0.001	1.000(1.000~1.000)
女方年龄	-0.036	0.009	15.803	＜0.001	0.964(0.947~0.982)
合并原发不孕	-0.335	0.089	14.193	0.002	0.715(0.601~0.851)
合并PCOS	1.884	0.142	175.570	＜0.001	6.580(4.979~8.694)
合并子宫内膜异位症	0.538	0.205	6.912	0.009	1.713(1.147~2.559)
注：变量赋值如下，FSH、LH、Gn时间、Gn剂量、女方年龄均以实际值赋值；合并原发不孕，是=1，否=0；合并PCOS，是=1，否=0；合并子宫内膜异位症，是=1，否=0。

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表 3 2PN+0PN组和2PN组接受ART治疗患者胚胎发育和临床结局比较

Table 3. Comparison of embryonic development and clinical outcomes in patients receiving ART between the 2PN+0PN group and the 2PN group

项目	2PN+0PN组	2PN组	统计量	P值
总周期数(个)	6 915	2 252
获卵数[M(P₂₅, P₇₅)，个]	12.0(7.0，18.0)	8.5 (5.0, 14.0)	-8.761^a	＜0.001
2PN受精数(个)	55 960	12 140
0PN受精数(个)	23 023	0
总受精率(2PN+0PN，%)	72.32(78 983/109 212)	63.52(12 140/19 111)	611.500^b	＜0.001
总卵裂率(2PN+0PN，%)	97.86(77 289/78 983)	98.01(11 899/12 140)	1.289^b	0.256
2PN卵裂率(%)	96.97(54 266/55 960)	100.00(11 899/11 899)	369.400^b	＜0.001
0PN卵裂率(%)	100.00(23 023/23 023)
优质胚胎率(%)	43.57(33 676/77 289)	47.98(5 709/11 899)	81.230^b	＜0.001
囊胚形成率(%)	55.01(42 518/77 289)	62.44(7 430/11 899)	231.100^b	＜0.001
优质囊胚率(%)	44.51(34 400/77 289)	48.15(5 729/11 899)	55.160^b	＜0.001
累计临床妊娠率(%)	79.68(5 510/6 915)	64.08(1 443/2 252)	225.800^b	＜0.001
累计活产率(%)	64.99(4 494/6 915)	54.97(1 238/2 252)	72.730^b	＜0.001
流产率(%)	9.38(517/5 510)	9.36(135/1 443)	0.001^b	0.975
新生儿体重(x±s，g)	3 231.0±651.1	3 196.0±658.4	1.642^c	0.101
Apgar评分(x±s，分)	9.95±0.33	9.93±0.37	1.858^c	0.063
注：^a为Z值，^b为χ²值，^c为t值。2PN受精数、0PN受精数反应总受精数的一部分，未进行比较。

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表 4 IVF周期中不同0PN衍生胚胎比例的胚胎发育及临床结果比较

Table 4. Comparison of embryo development and clinical outcomes of different 0PN embryo proportions in IVF cycles

项目	L组	M组	H组	统计量	P值
总周期数(个)	2 579	782	221
获卵数[M(P₂₅, P₇₅)，个]	15(10, 21)	10(5，16)^a	4(2, 9)^ab	327.800^c	＜0.001
受精率(%)	79.25(34 715/43 806)	77.72(7 957/10 238)^a	76.16(1 444/1 896)^a	20.206^d	＜0.001
卵裂率(%)	97.79(33 947/34 715)	98.74(7 857/7 957)^a	97.09(1 402/1 444)^b	34.544^d	＜0.001
优质胚胎率(%)	43.46(14 754/33 947)	46.47(3 651/7 857)^a	43.94(616/1 402)	23.404^d	＜0.001
囊胚形成率(%)	53.70(18 231/33 947)	57.46(4 515/7 857)^a	56.56(793/1 402)^a	38.911^d	＜0.001
优质囊胚率(%)	44.33(15 049/33 947)	47.36(3 721/7 857)^a	45.01(631/1 402)	23.654^d	＜0.001
累计临床妊娠率(%)	85.07(2 194/2 579)	78.26(612/782)^a	66.06(146/221)^ab	62.636^d	＜0.001
累计活产率(%)	70.61(1 821/2 579)	66.75(522/782)^a	55.20(122/221)^ab	24.498^d	＜0.001
流产率(%)	8.34(183/2 194)	5.39(33/612)^a	8.22(12/146)	6.373^d	0.041
新生儿体重(x±s，g)	3 223.0±625.0	3 190.0±654.4	3 365.0±680.5^ab	4.872^e	0.088
Apgar评分(x±s，分)	9.95±0.35	9.95±0.36	9.92±0.51	0.839^e	0.432
注：与L组比较，^aP＜0.05；与M组比较，^bP＜0.05。^c为Z值，^d为χ²值，^e为F值。

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表 5 0PN100%组与2PN100%组周期基本临床资料、胚胎发育及临床结局比较

Table 5. Comparison of basic clinical data, embryo development, and clinical outcome between the 0PN100% group and the 2PN100% group

项目	0PN100%组	2PN100%组	统计量	P值
总周期数(个)	72	100
FSH[M(P₂₅, P₇₅)，个]	8.950(6.640, 11.230)	9.065(7.158, 11.762)	-0.711^a	0.477
LH[M(P₂₅, P₇₅)，个]	4.400(2.990, 6.920)	4.710(3.380, 7.100)	-0.843^a	0.399
Gn时间(x±s，d)	9.77±3.09	10.16±2.12	0.806^b	0.422
Gn剂量[M(P₂₅, P₇₅)，IU]	2 287.50(1 743.75, 3 337.50)	2 250.00(1 725.00, 3 225.00)	0.142^a	0.887
女方年龄(x±s，岁)	33.56±6.48	33.21±6.27	0.329^b	0.743
原发不孕比例(%)	43.06(31/72)	45.00(45/100)	0.064^c	0.800
PCOS比例(%)	6.94(5/72)	8.00(8/100)	0.067^c	0.796
子宫内膜异位症比例(%)	1.39(1/72)	5.00(5/100)	1.621^c	0.203
获卵数[M(P₂₅, P₇₅)，个]	2(1，5)	11(8，13)	-8.891^a	＜0.001
受精率(%)	64.23(167/260)	64.62(579/896)	0.013^c	0.908
卵裂率(%)	100.00(167/167)	98.96(573/579)	1.745^c	0.187
优质胚胎率(%)	30.54(51/167)	53.40(306/573)	27.070^c	＜0.001
囊胚形成率(%)	55.09(92/167)	62.83(360/573)	3.257^c	0.196
优质囊胚率(%)	32.34(54/167)	53.05(304/573)	22.230^c	＜0.001
累计临床妊娠率(%)	40.28(29/72)	67.00(67/100)	12.120^c	0.001
累计活产率(%)	30.56(22/72)	60.00(60/100)	14.550^c	＜0.001
流产率(%)	3.45(1/29)	11.94(8/67)	1.718^c	0.424
新生儿体重(x±s，g)	3 137.0±643.5	3 101.0±538.0	0.993^b	0.321
Apgar评分(x±s，分)	10.00±0.00	10.00±0.06	1.416^b	0.157
注：^a为Z值，^b为t值，^c为χ²值。

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参考文献(18)

[1]	程静娴, 王春艳, 王婕妤, 等. 辅助生殖夫妇婚姻状况的影响因素及其对妊娠结局的影响[J]. 中华全科医学, 2023, 21(5): 744-748. doi: 10.16766/j.cnki.issn.1674-4152.002973 CHENG J X, WANG C Y, WANG J Y, et al. Factors influencing marital status in assisted reproduction couples and its impact on pregnancy outcomes[J]. Chinese Journal of General Practice, 2023, 21(5): 744-748. doi: 10.16766/j.cnki.issn.1674-4152.002973
[2]	VANDER BORGHT M, WYNS C. Fertility and infertility: definition and epidemiology[J]. Clin Biochem, 2018, 62: 2-10. doi: 10.1016/j.clinbiochem.2018.03.012
[3]	吴娴, 张爱军. 辅助生殖实验室技术的改进[J]. 实用妇产科杂志, 2020, 36(4): 249-251. WU X, ZHANG A J. Improvements in laboratory techniques for assisted reproduction[J]. Journal of Practical Obstetrics and Gynecology, 2020, 36(4): 249-251.
[4]	KEMPER J M, LIU Y H, AFNAN M, et al. What happens to abnormally fertilized embryos? A scoping review[J]. Reprod Biomed Online, 2023, 46(5): 802-807. doi: 10.1016/j.rbmo.2023.02.005
[5]	赵双丹, 赵志明, 赵兵依, 等. 常规体外受精后未见原核胚胎的相关因素分析及临床价值探讨[J]. 中国实用妇科与产科杂志, 2021, 37(2): 223-227. ZHAO S D, ZHAO Z M, ZHAO B Y, et al. Analysis of the factors related to the non-pronuclear zygotes embryos after conventional in vitro fertilization and the clinical value[J]. Chinese Journal of Practical Gynecology and Obstetrics, 2021, 37(2): 223-227.
[6]	ZHAO H B, LIU H, LI M, et al. Clinical outcomes following frozen-thawed blastocyst transfers with blastocysts derived from different cell numbers on day 3: a retrospective cohort study[J]. J Assist Reprod Genet, 2020, 37(3): 641-648. doi: 10.1007/s10815-019-01664-x
[7]	DAI X L, GAO T T, XIA X Y, et al. Analysis of biochemical and clinical pregnancy loss between frozen-thawed embryo transfer of blastocysts and day 3 cleavage embryos in young women: a comprehensive comparison[J]. Front Endocrinol, 2021, 127: 85658. DOI: 10.3389/fendo.2021.785658.
[8]	EZOE K, TAKAHASHI T, SHIMAZAKI K, et al. Human 1PN and 3PN zygotes recapitulate all morphokinetic events of normal fertilization but reveal novel developmental errors[J]. Hum Reprod, 2022, 37(10): 2307-2319. doi: 10.1093/humrep/deac177
[9]	PAZ M V, CHIERA M, HOVANYECZ P, et al. Blastocysts derived from 0PN oocytes: genetic and clinical results[J]. JBRA Assist Reprod, 2020, 24(2): 143-146.
[10]	李澎涛, 殷晨星, 孟娜娜, 等. 0PN来源胚胎形成的影响因素及其利用价值分析[J]. 生殖医学杂志, 2023, 32(10): 1503-1509. LI P T, YIN C X, MENG N N, et al. Analysis of influencing factors and utilization value of 0PN-derived embryo formation[J]. Journal of Reproductive Medicine, 2023, 32(10): 1503-1509.
[11]	ZHU J, WANG C L, CAO Z Y, et al. Developmental competence and neonatal outcomes of nonpronuclear zygotes following single vitrified-warmed blastocyst transfers using propensity score matching analysis[J]. Arch Gynecol Obstet, 2024, 309(1): 295-304.
[12]	张旸, 翟妍, 付磊, 等. 异常原核数来源胚胎的妊娠结局分析[J]. 中华医学杂志, 2019, 99(29): 2308-2310. ZHANG Y, ZHAI Y, FU L, et al. Analysis of pregnancy outcome of embryos derived from abnormal prokaryotes[J]. National Medical Journal of China, 2019, 99(29): 2308-2310.
[13]	KOBAYASHI T, ISHIKAWA H, ISHII K, et al. Time-lapse monitoring of fertilized human oocytes focused on the incidence of 0PN embryos in conventional in vitro fertilization cycles[J]. Sci Rep, 2021, 11(1): 18862. DOI: 10.1038/s41598-021-98312-1.
[14]	XIE P Y, TANG Y, HU L, et al. Identification of biparental and diploid blastocysts from monopronuclear zygotes with the use of a single-nucleotide polymorphism array[J]. Fertil Steril, 2018, 110(3): 545-554. doi: 10.1016/j.fertnstert.2018.04.034
[15]	FU L, ZHOU W H, LI Y. Development and frozen-thawed transfer of non-pronuclear zygotes-derived embryos in IVF cycles[J]. Eur J Obstet Gynecol Reprod Biol, 2021, 264: 206-211. doi: 10.1016/j.ejogrb.2021.07.033
[16]	石小丹, 王云, 赵楠, 等. 着床前遗传学筛查周期中不同受精来源囊胚发育速度与染色体整倍性分析[J]. 实用医技杂志, 2019, 26(10): 1225-1227. SHI X D, WANG Y, ZHAO N, et al. Analysis of the development rate and chromosome ploidy of blastocyst from different sources of fertilization in the preimplantation genetic testing cycle[J]. Journal of Practical Medical Techniques, 2019, 26(10): 1225-1227.
[17]	TONG X M, JIN J M, XUE Y M, et al. Clinical outcomes of frozen-thawed blastocysts from zygotes with no or one pronucleus for in vitro fertilization and intracytoplasmic sperm injection cycles[J]. Arch Gynecol Obstet, 2023, 308(3): 1015-1022. doi: 10.1007/s00404-023-07118-1
[18]	FRAGOULI E, MUNNE S, WELLS D. The cytogenetic constitution of human blastocysts: insights from comprehensive chromosome screening strategies[J]. Hum Reprod Update, 2019, 25(1): 15-33. doi: 10.1093/humupd/dmy036