Our objective was to determine the effects of methionine and choline supplementation during the pre- and postpartum periods on preimplantation embryos of Holstein cows. Multiparous cows were assigned in a randomized complete block design into four treatments from 21 d before calving to 30 DIM. Treatments (TRT) were: CON (n=8, fed the close-up and fresh cow diets with a Lys:Met=3.5:1), MET (n=9, fed the basal diet + methionine, Smartamine® M to a Lys:Met=2.9:1), CHO (n=8, fed the basal diets + choline 60 g/d, Reashure®), and MIX (n=11, fed the basal diets plus Smartamine M® to a Lys:Met=2.9:1 and 60 g/d Reashure®). From 30 ± 1 to 72 ± 1 DIM the cows were randomly assigned to two groups (GRP); control (CTL; n=16, fed a basal diet with a 3.5:1 Lys:Met) and methionine (SMT; n=20, fed the basal diet + methionine to 2.9:1 Lys:Met). On d 60, dominant follicles greater than 5 mm were aspirated using an ultrasound-guided transvaginal approach. A CIDR® device was inserted in all cows after follicular aspiration and superovulation began at d 61.5 using FSH treatment equivalent to 400 mg of NIHFSHP1 (Folltropin®) in 8 decreasing doses at 12 h intervals over a 4 d period. During the superovulatory period, all cows received two PGF2α injections at d 63 and d 64 (concomitant with the 5th and 7th FSH injections), and CIDR was withdrawn at d 65. Twenty-four hours after CIDR withdrawal, ovulation was induced with GnRH. Cows were artificially inseminated at 12 h and 24 h after GnRH. Embryos were flushed 6.5 d after artificial insemination. Global methylation of the embryos was assessed by immunofluorescent labeling with 5-methylcytosine, while lipid content was assessed by staining with Nile Red. Nuclear staining (propidium iodide or Hoescht 33342) was used to count the total number of cells/embryo. Statistical analysis was performed using the MIXED procedure of SAS. Methylation of the DNA did was not different (P>0.05) among treatments but there was a TRT × GRP interaction (P=0.03). Embryos from cows in CON (– 21 to 72 d) had greater (P = 0.04) methylation (0.87 ± 0.09) than the embryos from cows in MET and CTL (0.44 ± 0.07).Embryos from cows in SMT had greater lipid content (P=0.04; 7.02 ± 1.03) than CTL (3.61 ± 1.20). There was not difference (P>0.05) for cells/embryo, embryo recovery rate per flushing, number of embryos recovered, embryo quality, embryo stage, and numbers of CL at flushing (CTL: 61.48±5.12, 0.69 %, 7.69±1.39, 1.63±0.25, 3.92±0.12, 9.33±1.18; and SMT: 54.92±3.9, 0.80 %, 8.72±1.18, 1.99±0.20, 4.13±0.10, 11.47±0.99, respectively). In conclusion, supplementation of methionine, choline or both methionine and choline affect embryo methylation and lipid content.