Colorado Center for Reproductive Medicine, Lone Tree, CO, USA.
J Assist Reprod Genet. 2020 Apr;37(4):747-752. doi: 10.1007/s10815-020-01714-9. Epub 2020 Feb 18.
To establish parameters during mouse extended embryo culture that accurately predict fetal developmental potential of a blastocyst without performing embryo transfer.
Embryos of three varying qualities were produced: poor quality embryos produced from in vitro matured oocytes (IVM), intermediate quality embryos produced from in vivo matured oocytes followed by in vitro fertilization and embryo culture (IVF); high quality embryos developed in vivo (VIVO). Embryonic day (E) 3.5 embryos from each group with similar morphologies were used for surgical embryo transfer to assess implantation and fetal developmental potential, in addition to placing these embryos into extended culture until E 8.5 to examine outgrowth area, egg cylinder volume, epiblast cell number, and outgrowth morphologies by immunofluorescence and 3D confocal microscopy.
The proportional differences in epiblast cell number are strikingly similar to fetal development following embryo transfer, suggesting that this parameter may be indicative of the potential of an embryo to successfully develop into a fetus.
Extended embryo culture provides more accurate information regarding developmental potential than blastocyst morphological assessment. Specifically, epiblast cell number is an accurate and valuable predictor of fetal developmental potential. This work sets the stage for routine evaluation of embryo quality past the time embryos would normally be transferred. The ability to determine post implantation potential without embryo transfer may greatly improve efforts to culture higher quality embryos in vitro for human IVF, as well as reducing animal use and eliminating confounding maternal factors associated with embryo transfer experiments in research.
建立小鼠胚胎延长培养过程中的参数,在不进行胚胎移植的情况下准确预测囊胚的胎儿发育潜能。
产生了三种不同质量的胚胎:体外成熟卵母细胞(IVM)产生的质量差的胚胎、体内成熟卵母细胞受精和胚胎培养(IVF)产生的中等质量胚胎;体内发育的高质量胚胎(VIVO)。从每个组中选择具有相似形态的 E3.5 胚胎进行手术胚胎移植,以评估植入和胎儿发育潜能,此外还将这些胚胎放入延长培养中,直到 E8.5 以通过免疫荧光和 3D 共聚焦显微镜检查外胚层细胞数量、囊胚体积、上胚层细胞数量和外胚层细胞形态。
上胚层细胞数量的比例差异与胚胎移植后的胎儿发育非常相似,表明该参数可能表明胚胎成功发育为胎儿的潜力。
胚胎延长培养提供的关于发育潜能的信息比囊胚形态评估更准确。具体来说,上胚层细胞数量是胎儿发育潜能的准确而有价值的预测指标。这项工作为常规评估胚胎质量奠定了基础,超过了胚胎通常转移的时间。无需胚胎移植即可确定植入后潜力的能力,可能会极大地改善体外培养高质量胚胎进行人类体外受精的努力,同时减少动物使用并消除与胚胎转移实验相关的母体因素。