Differential impacts of gonadotrophins, IVF and embryo culture on mouse blastocyst development

Published:March 12, 2019DOI:


      Research question

      Conception via assisted reproductive technology (ART) increases the risk of type 2 diabetes and cardiovascular disease in adulthood. Underlying differences between ART-conceived and in-vivo-conceived embryos that contribute to this increased risk are, however, not known.


      This study examined the developmental characteristics of mouse blastocysts derived from ART- compared with in-vivo-conceived embryos. To determine the effect of ovarian stimulation versus IVF versus in-vitro embryo culture on phenotype, six distinct groups of blastocysts were generated. Female mice were naturally cycling or treated with high or mild doses of gonadotrophin, followed by natural mating or IVF under clinical conditions. Embryo morphokinetics were assessed by continuous time-lapse monitoring. Cell lineage allocation to the inner cell mass (Oct4+) or trophectoderm (Cdx2+) was determined by immunohistochemistry, and mitochondrial DNA (mtDNA) copy number was measured by quantitative PCR.


      Ovarian stimulation increased embryo number but reduced the percentage of blastocysts. Morphokinetic analysis showed that gonadotrophin treatment led to advanced development (P < 0.05) due to earlier post-pronuclear breakdown. The blastocyst rate was reduced in IVF embryos compared with those fertilized in vivo before culture (P < 0.001). Morphokinetics showed that embryo development was slower in all the IVF groups (P < <0.05), due to a delay from the 3-cell stage. A reduced total and trophectoderm cell number was observed in all groups of cultured blastocysts compared with naturally conceived blastocysts (P < 0.01). Gonadotrophin treatment did not affect the blastocyst mtDNA copy number; however, IVF embryos exhibited reduced mtDNA copy number compared with naturally conceived embryos.


      Ovarian stimulation, IVF and in-vitro culture differentially impair blastocyst developmental kinetics, differentiation and mtDNA copy number.


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      Miaoxin Chen is a specialist physician and an Associate Professor of Tongji University in China. He completed his PhD and postdoctoral research at the University of Adelaide in Australia from October 2010 to January 2016, and his thesis was awarded the Dean's Commendation for Doctoral Thesis Excellence in 2014. His research interests are the effects of assisted reproductive technologies on embryos and long term health in humans and mice.
      Key message
      Using a mouse model to isolate specific ART processes, the study showed that culture environment, particularly at fertilization, influenced key markers of embryo development, cell number and mitochondrial DNA. These findings have implications for understanding the basis of embryo phenotypes that may contribute to the increased risk of metabolic diseases in ART-conceived individuals.