Lipotropes (methionine, choline, folate, and vitamin B12) play a key role in one-carbon metabolism. We hypothesized that maternal methyl (lipotropic) nutrients would induce epigenetic modification that influences mammary development and carcinogenesis from dam to offspring (F1 & F2). Pregnant Sprague-Dawley rats were fed either control or lipotropes diet until parturition. Global DNA methylation significantly increased in the lipotropes group of both generations. Histone deacetylase 1 (Hdac1) increased in F2 in mammary tissues. In the F2 mammary tumor tissues, tumor incidence, and multiplicity were lowered by maternal methyl nutrients. Latency period was longer and tumor volumes decreased in the lipotropes group. Methyl nutrients significantly decreased transcription of histone deacetylase 1 (Hdac1) and protein expression in the lipotropes group of F2 mammary tumor tissues (P=0.02). Data suggest that maternal dietary lipotropes may be involved in permanent epigenetic alteration of gene expression, reducing the risk of mammary cancer in subsequent generations, including F2 generation.