In Vitro Fertilization Induces Premature Reproductive Aging and Multigenerational Epigenetic Changes in Female Mouse Offspring

In vitro fertilization (IVF) is an assisted reproductive technology that has enabled millions of births. Although considered safe, emerging evidence suggests that IVF may have long-term health adverse outcomes in offspring, though its impact on female reproduction, especially on reproductive aging is largely unknown. Here, we use a mouse model to investigate how IVF influences female reproductive and transgenerational health outcomes. We assessed IVF-conceived fetal (E18.5) and adult (12-week and 39-week) female offspring, identifying alterations in ovarian-to-body weight ratios, ovarian morphology, serum sex hormone levels, and transcriptomic and DNA methylation profiles in ovaries, oocytes, and cumulus cells, consistent with biomarkers of accelerated reproductive aging. The offspring of IVF females and wild-type males exhibited altered fetal-to-placental weight ratios, placental morphology, and dysregulated placental gene expression. At 12-weeks-of-age, offspring showed increased body weight, disrupted lipid and glucose metabolism, and transcriptomic and epigenetic changes in liver and reproductive organs. Our results highlight the need for deeper mechanistic understanding of assisted reproduction's long-term and multigenerational impacts on female reproductive health. Overall design: Gene expression analysis using Total RNA extracted from F1: E18.5 ovaries (n=10, Natural=4 and IVF=6), 12 weeks ovaries (n=10, Natural=5 and IVF=5), 39 weeks ovaries (n=10, Natural=4 and IVF=6), 12 weeks single oocytes (n=20, Natural=10 and IVF=10), 39 weeks single oocytes (n=20, Natural=10 and IVF=10), 12 weeks cumulus cells (n=4, Natural=2 and IVF=2), 39 weeks cumulus cells (n=4, Natural=2 and IVF=2) and embryonic day 13.5 female gonads (n=9, Natural=4 and IVF=5). F2: 12 weeks testis (n=12, Natural=5 and IVF=7), 12 weeks male livers (n=10, Natural=5 and IVF=5), 12 weeks ovaries (n=9, Natural=4 and IVF=5), 12 weeks female livers (n=10, Natural=4 and IVF=4), using either Illumina NovaSeq6000 or NextSeq1000.