The duration of embryo culture after mouse IVF differentially affects cardiovascular and metabolic health in male offspring.
The duration of embryo culture after mouse IVF differentially affects cardiovascular and metabolic health in male offspring.
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DOI：
10.1093/humrep/deaa205
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发表时间：
2020-11-01
期刊：
影响因子：
--
通讯作者：
Fleming TP
中科院分区：
文献类型：
--
作者：
Aljahdali A;Airina RKRI;Velazquez MA;Sheth B;Wallen K;Osmond C;Watkins AJ;Eckert JJ;Smyth NR;Fleming TP
IVF后的长期健康结果是否取决于移植前胚胎培养的持续时间？使用小鼠模型，我们证明，在男性而不是女性后代，不良心血管（CV）健康更可能与长期培养到囊胚阶段，但代谢功能障碍更可能，如果胚胎移植（ET）发生在早期卵裂阶段。ART与后代不良CV和代谢健康的风险增加有关，这些发现已在没有父母不育问题的动物模型中得到证实。目前尚不清楚哪些特定的ART治疗可能导致这些风险。在临床ART中越来越多地使用囊胚移植，而不是卵裂期移植，这似乎不会损害出生儿童的围产期健康，但长期健康影响尚不清楚。产生五个小鼠组，包括：（i）自然交配（NM）-自然交配、非超排卵和未受干扰的妊娠;（ii）IV-ET-2细胞-体内衍生的两细胞胚胎，其收集自超排卵的母亲，立即ET给受体;（iii）IVF-ET-2细胞-IVF产生的胚胎，其来自超排卵母亲的卵母细胞，在ET给受体之前培养至两细胞阶段;（iv）IV-ET-BL-体内衍生的胚泡收集自超数排卵的母亲，立即ET给受体;（v）IVF-ET-BL-IVF产生的胚胎，来自超数排卵的母亲的卵母细胞，在ET给受体之前培养至胚泡阶段。分析了雄性和雌性后代的生长、CV和健康代谢标志物。每组产生8-13窝用于分析;通过多水平随机效应回归分析产后数据，以考虑母亲间和母亲内变异和窝仔数。C57/BL 6雌性小鼠（3-4周龄）用于卵母细胞生产; CBA雄性小鼠用于精子和人输卵管液培养基用于IVF。胚胎移植前，将胚胎在2细胞期移植到MF 1假孕受体或在富含钾的合成输卵管培养基中培养至囊胚期。收集来自C57 BL 6 × CBA交配的对照体内胚胎，并在二细胞或囊胚期立即移植。出生后测定包括27周的生长率; 9、15和21周的收缩压（SBP）;剔除时（27周）的肺和血清血管紧张素转换酶（ACE）活性;葡萄糖耐量试验（GTT; 27周）;基础葡萄糖和胰岛素水平（27周）;以及使用油红O成像的肝脏冷冻切片中的脂质蓄积（27周）。体外受精形成的囊胚发育速度慢于体外培养形成的囊胚，囊胚细胞数少于体外培养形成的囊胚（P < 0.05）。与NM组相比，4个试验处理组的仔鼠出生后生长速度均有所提高（P < 0.05）。IVF-ET-BL组SBP、血清和肺ACE及心/体重均高于IVF-ET-2细胞组（P < 0.05），且高于其他治疗组，SBP与肺ACE呈正相关（P < 0.05）。IVF-ET-2Cell组男性的葡萄糖代谢能力（GTT AUC）较IVF-ET-BL组差，基础胰岛素水平较高（P < 0.05）。IVF-ET-2Cell组男性的葡萄糖/胰岛素比值与体重呈显著负相关（P < 0.05）。IVF-ET-2细胞组的肝重/体重、肝脂滴直径和密度均高于IVF-ET-BL组（P < 0.05）。IVF组的健康特征比体内对照组差，表明结果不是由背景技术（超排卵，ET）引起的。在雌性后代中未发现培养持续时间对健康的一致影响。N/A.实验动物模型的结果不能外推到人类。然而，他们是有价值的概念模型，在这种情况下，在没有混淆父母不育，在评估ART操作的安全性。该研究表明，与IVF和较短的卵裂期ET相比，IVF后胚胎培养至ET前囊胚的持续时间较长，导致男性CV健康功能障碍增加。然而，雄性后代的代谢健康状况较差后较短的培养时间比较长的。这种区别表明ART后CV和代谢健康表型的起源可能不同。卵裂期ET后男性的代谢健康状况较差，这与ET时发生的胚胎基因组激活相一致。这项工作得到了欧盟FP 7-CP-FP Epihealth计划（278418）和FP 7-PEOPLE-2012-ITN EpiHealthNet计划（317146）的支持，生物技术和生物科学研究理事会（BBSRC）（BB/F007450/1），以及沙特政府、吉达大学和阿卜杜勒-阿齐兹国王大学，作者没有利益冲突需要声明。
Do the long-term health outcomes following IVF differ depending upon the duration of embryo culture before transfer? Using a mouse model, we demonstrate that in male but not female offspring, adverse cardiovascular (CV) health was more likely with prolonged culture to the blastocyst stage, but metabolic dysfunction was more likely if embryo transfer (ET) occurred at the early cleavage stage. ART associate with increased risk of adverse CV and metabolic health in offspring, and these findings have been confirmed in animal models in the absence of parental infertility issues. It is unclear which specific ART treatments may cause these risks. There is increasing use of blastocyst, versus cleavage-stage, transfer in clinical ART which does not appear to impair perinatal health of children born, but the longer-term health implications are unknown. Five mouse groups were generated comprising: (i) natural mating (NM)—naturally mated, non-superovulated and undisturbed gestation; (ii) IV-ET-2Cell—in-vivo derived two-cell embryos collected from superovulated mothers, with immediate ET to recipients; (iii) IVF-ET-2Cell—IVF generated embryos, from oocytes from superovulated mothers, cultured to the two-cell stage before ET to recipients; (iv) IV-ET-BL—in-vivo derived blastocysts collected from superovulated mothers, with immediate ET to recipients; (v) IVF-ET-BL—IVF generated embryos, from oocytes from superovulated mothers, cultured to the blastocyst stage before ET to recipients. Both male and female offspring were analysed for growth, CV and metabolic markers of health. There were 8–13 litters generated for each group for analyses; postnatal data were analysed by multilevel random effects regression to take account of between-mother and within-mother variation and litter size. C57/BL6 female mice (3–4 weeks old) were used for oocyte production; CBA males for sperm with human tubal fluid medium were used for IVF. Embryos were transferred (ET) to MF1 pseudo-pregnant recipients at the two-cell stage or cultured in synthetic oviductal medium enriched with potassium medium to the blastocyst stage before ET. Control in-vivo embryos from C57BL6 × CBA matings were collected and immediately transferred at the two-cell or blastocyst stage. Postnatal assays included growth rate up to 27 weeks; systolic blood pressure (SBP) at 9, 15 and 21 weeks; lung and serum angiotensin-converting enzyme (ACE) activity at time of cull (27 weeks); glucose tolerance test (GTT; 27 weeks); basal glucose and insulin levels (27 weeks); and lipid accumulation in liver cryosections using Oil Red O imaging (27 weeks). Blastocysts formed by IVF developed at a slower rate and comprised fewer cells that in-vivo generated blastocysts without culture (P < 0.05). Postnatal growth rate was increased in all four experimental treatments compared with NM group (P < 0.05). SBP, serum and lung ACE and heart/body weight were higher in IVF-ET-BL versus IVF-ET-2Cell males (P < 0.05) and higher than in other treatment groups, with SBP and lung ACE positively correlated (P < 0.05). Glucose handling (GTT AUC) was poorer and basal insulin levels were higher in IVF-ET-2Cell males than in IVF-ET-BL (P < 0.05) with the glucose:insulin ratio more negatively correlated with body weight in IVF-ET-2Cell males than in other groups. Liver/body weight and liver lipid droplet diameter and density in IVF-ET-2Cell males were higher than in IVF-ET-BL males (P < 0.05). IVF groups had poorer health characteristics than their in-vivo control groups, indicating that outcomes were not caused specifically by background techniques (superovulation, ET). No consistent health effects from duration of culture were identified in female offspring. N/A. Results from experimental animal models cannot be extrapolated to humans. Nevertheless, they are valuable to develop conceptual models, in this case, in the absence of confounding parental infertility, in assessing the safety of ART manipulations. The study indicates that longer duration of embryo culture after IVF up to blastocyst before ET leads to increased dysfunction of CV health in males compared with IVF and shorter cleavage-stage ET. However, the metabolic health of male offspring was poorer after shorter versus longer culture duration. This distinction indicates that the origin of CV and metabolic health phenotypes after ART may be different. The poorer metabolic health of males after cleavage-stage ET coincides with embryonic genome activation occurring at the time of ET. This work was supported through the European Union FP7-CP-FP Epihealth programme (278418) and FP7-PEOPLE-2012-ITN EpiHealthNet programme (317146) to T.P.F., the Biotechnology and Biological Sciences Research Council (BBSRC) (BB/F007450/1) to T.P.F., and the Saudi government, University of Jeddah and King Abdulaziz University to A.A. The authors have no conflicts of interest to declare.
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