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Objectives: In this observational study, we examined whether maternal methyl nutrient intake or status was associated with placental and cord blood DNA methylation in GDM and non-GDM pregnancies.

Methods: We recruited pregnant women with and without GDM during gestational week 25–32 and assessed their methyl nutrient intake and blood status. We collected placenta and cord blood at delivery.

Results: Our results demonstrated that maternal betaine intake was associated with lower cord blood IGF2 methylation, while maternal choline intake was associated with higher cord blood CRH methylation. These associations may have functional implications as demonstrated by lower birth weight related to higher maternal betaine intake and lower cord blood stress hormone cortisol levels with higher maternal choline intake. Maternal folate status was associated with lower placental IGF2 DNA methylation, which may be related to the lower DNA methyltransferase expression with high folate status. GDM status did not modify most of the diet and epigenetic interactions. Conclusions: In conclusion, maternal intake of methyl nutrients has a complex relationship with fetal DNA methylation levels regardless of GDM.