Long-term effect of in vitro culture of mouse embryos with serum on mRNA expression of imprinting genes, development, and behavior

Abstract

The long-term developmental and behavioral consequences of mammalian embryo culture are unknown. By altering the culture medium with the addition of FCS, we wanted to determine whether mouse embryos cultured under suboptimal conditions develop aberrant mRNA expression of imprinting genes at the blastocyst stage and whether fetal development, growth, and behavior of adult mice are affected. One-cell embryos obtained from superovulated female B6CBAF1 mice were cultured for 4 days in K ؉ -modified simplex optimized medium in the presence of either 10% FCS or 1 g͞liter BSA. After embryo transfer, born animals were submitted to several developmental and behavior tests. The mRNA expression of some imprinting genes was significantly affected in blastocysts cultured in the presence of FCS. Two of the eight measures of preweaning development and some specific measures of neuromotor development, such as the walking activity, were delayed in the group originated with FCS. After 34 weeks, the weight of female mice cultured in vitro in the presence of FCS was significantly higher than controls. In addition, the locomotion activity of mice was altered at 5 and 15 months. Anatomopathological and histological analysis of animals at 20 months of age showed some large organs and an increase in pathologies. We have found that mice derived from embryos cultured with FCS exhibited specific behavioral alterations in anxiety and displayed deficiencies in implicit memories. Our data indicate that long-term programming of postnatal development, growth, and physiology can be affected irreversibly during the preimplantation period of embryo development by suboptimal in vitro culture.embryo culture ͉ organ weight ͉ postnatal defects

Differences in developmental competence and gene expression profiles between buffalo (Bubalus bubalis) preimplantation embryos cultured in three different embryo culture media

“…In this study we analyzed expression of a panel of developmentally important genes in IVF embryos of different developmental stages cultured in undefined and defined media. Our results support the hypothesis that in vitro conditions have an effect on patterns of embryonic gene expression in buffalo, as reported in other species including bovine and mouse (Fernández-Gonzalez et al 2004;Schwarzer et al 2012;Arias et al 2013). Furthermore, the study by Schwarzer et al (2012), found that 13 different embryo culture protocols resulted in distinct cellular and molecular phenotypes in mouse embryos.…”

Genomic landscape of human allele-specific DNA methylation

“…A substantial part may also be due to epigenotypic variation between the cells: Those from culture will exhibit the associated epigenomic effects. It has been known for some time that permanent cell lines have altered DNA methylation (41,42), and more recently it was shown that aberrant methylation is correlated with passage number (43). We also observed some examples which might indicate an effect of culture conditions, such as the MEG3 AMRs (Fig.…”

Serum free embryo culture medium improves in vitro survival of bovine blastocysts to vitrification

“…Despite the efficiency of in vitro production systems in yielding considerable numbers of embryos, the major fall-off in in vitro embryo development occurs between the two-cell and blastocyst stages, suggesting that postfertilization embryo culture is the most critical period of the process in terms of determining the blastocyst quality [35,36].…”