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个体文化和培养过程中的大气氧含量会影响小鼠胚胎植入前的代谢和植入后的发育。

Individual culture and atmospheric oxygen during culture affect mouse preimplantation embryo metabolism and post-implantation development.

机构信息

School of BioSciences, The University of Melbourne, Parkville Victoria 3010, Australia.

School of BioSciences, The University of Melbourne, Parkville Victoria 3010, Australia.

出版信息

Reprod Biomed Online. 2019 Jul;39(1):3-18. doi: 10.1016/j.rbmo.2019.03.102. Epub 2019 Mar 11.

Abstract

RESEARCH QUESTION

Does single embryo culture under atmospheric or reduced oxygen alter preimplantation metabolism and post-implantation development compared with culture in groups?

DESIGN

Mouse embryos were cultured under 5% or 20% oxygen, individually or in groups of 10. Spent media were analysed after 48, 72 and 96 h of culture. Blastocysts were assessed by outgrowth assay or transferred to pseudo-pregnant recipients, and fetal and placental weight, length and morphology were assessed.

RESULTS

Compared with group culture, individually cultured blastocysts had lower net consumption of glucose and aspartate and higher glutamate production. Atmospheric oxygen reduced uptake of glucose and aspartate and increased production of glutamate and ornithine compared with 5% oxygen. Combining 20% oxygen and single culture resulted in further metabolic changes: decreased leucine, methionine and threonine consumption. Under 5% oxygen, individual culture decreased placental labyrinth area but had no other effects on fetal and placental development or outgrowth size compared with group culture. Under 20% oxygen, however, individual culture reduced outgrowth size and fetal and placental weight compared with group-cultured embryos.

CONCLUSIONS

Preimplantation metabolism of glucose and amino acids is altered by both oxygen and individual culture, and fetal weight is reduced by individual culture under atmospheric oxygen but not 5% oxygen. This study raises concerns regarding the increasing prevalence of single embryo culture in human IVF and adds to the existing evidence regarding the detrimental effects of atmospheric oxygen during embryo culture. Furthermore, these data demonstrate the cumulative nature of stress during embryo culture and highlight the importance of optimizing each element of the culture system.

摘要

研究问题

与分组培养相比,大气或低氧条件下的单个胚胎培养是否会改变胚胎植入前的代谢和胚胎植入后的发育?

设计

将小鼠胚胎分别在 5%或 20%的氧气中进行培养,或者以 10 个胚胎为一组进行培养。在培养 48、72 和 96 小时后分析废弃培养基。通过外胚层发育测定或移植到假孕受体中来评估囊胚,并评估胎儿和胎盘的重量、长度和形态。

结果

与分组培养相比,单个培养的囊胚葡萄糖和天冬氨酸的净消耗减少,谷氨酸的产生增加。与 5%氧气相比,大气氧气降低了葡萄糖和天冬氨酸的摄取,并增加了谷氨酸和鸟氨酸的产生。与 20%氧气和单个培养相结合,导致进一步的代谢变化:减少亮氨酸、蛋氨酸和苏氨酸的消耗。在 5%氧气下,与分组培养相比,单个培养减少了胎盘绒毛区面积,但对胎儿和胎盘发育或外胚层大小没有其他影响。然而,在 20%氧气下,与分组培养的胚胎相比,单个培养减少了外胚层发育和胎儿及胎盘重量。

结论

氧气和单个培养都会改变胚胎植入前葡萄糖和氨基酸的代谢,大气氧气下的单个培养会降低胎儿体重,但 5%氧气下则不会。本研究对人类体外受精中日益流行的单个胚胎培养提出了担忧,并增加了胚胎培养过程中大气氧气的有害影响的现有证据。此外,这些数据表明胚胎培养过程中应激的累积性质,并强调了优化培养系统每个元素的重要性。

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