Assessment of the Long-Term and Transgenerational Consequences of Perturbing Preimplantation Embryo Development in Mice1

Abstract

Perturbations of the development of preimplantation embryos may have long-term consequences for the health of progeny. There are no standardized methods for assessing such risks. The OECD/OCDE 416 Guideline for Testing of Chemicals (Two-Generation Reproduction Toxicity Study) is a standardized assay for detecting potential toxic effects of chemicals. The present study assessed the utility of this guideline for identifying long-term consequences of perturbing preimplantation development. Extended culturing of mammalian zygotes commonly results in retarded preimplantation development. Mouse zygotes were cultured in vitro for 96 h until the blastocyst stage (cultured blastocysts) or blastocysts were collected from the Day-3.5 uterus (in vivo blastocysts). The resulting blastocysts were transferred to the uteri of pseudopregnant recipients (P generation). Progeny from both treatments were mated for a further two generations (F1 and F2 generations). There was no effect of treatment group on gross fertility across the generations tested. Progeny of the cultured blastocysts had lower body weights to the time of weaning compared to in vivo blastocysts in the P and F1 generations, but not in the F2 generation. At maturity, there was no effect of treatment group on body weight, although thyroid weight was higher in the in vivo blastocyst group in the P generation, while the brain, pituitary, and kidneys were larger in the progeny of the cultured blastocysts of the F1 generation. The OECD/OCDE 416 assessment may have a role as a standardized test for the assessment of the biological consequences of perturbing the growth environment of the preimplantation embryo. Embryo culture influenced the somatometric parameters of the resulting progeny, some of which were maintained across a generation.

Embryo vitrification in rabbits: Consequences for progeny growth

“…In addition, several authors re-ported modifications in adult organs size due to embryo manipulation. For instance, Mahsoudi et al [45] and Calle et al [46] observed that manipulation of preimplantation mouse embryos resulted in altered organ sizing, and Serrano et al [47] observed that modifications in embryo culture conditions modify the adult phenotypes, increasing the sensitivity to development of liver steatosis in females. These phenotypes could be the consequence of messenger RNA expression differences of metabolic relevant genes due to changes in methylation pattern [48,49].…”

Peroxides in mineral oil used for in vitro fertilization: defining limits of standard quality control assays

“…The data in this study demonstrated a dose dependent effect of CH on blastocyst cell number at concentrations of peroxide that pass both the 1-cell and 2-cell group culture MEA. Since mice derived from in vitro blastocyst culture have decreased birth weight and weaning weight when compared with in vivo conceived offspring [23], this effect could be due to undetected sub-lethal doses of toxins present in the culture environment. These results highlight the need for more sensitive quality control assays.…”

Considerations Regarding Embryo Culture Conditions: From Media to Epigenetics

“…One of the main external factors considered to being capable of altering embryo development and affecting offspring health is the oxygen tension during culture (21). There are various animal studies suggesting that altered gene expression may take place and is majorly dependent on the IVF environment and culture media (86)(87)(88)(89)(90). Applying these data to humans is challenging and requires careful extrapolation.…”