DNA methylation/demethylation and histone modifications represent classic interdependent epigenetic mechanisms controlling gene expression. Di-methylation of lysine 4 on histone 3 (H3K4me2) imprinting and DNA demethylase Tet methylcytosine dioxygenase- 2 (Tet-2) binding at lineage-specific gene loci has been proposed as the central epigenetic mechanisms controlling retention of lineage identity and cellular memory of vascular smooth muscle cells (SMCs) during SMC de-differentiation. This study aimed to assess whether similar histone modifications and Tet-2 binding occur at SMC loci in both stem cell-derived SMC-like cells (myogenic progeny) and non-SMCs (adipogenic, osteogenic and vasculogenic progeny) typical of arteriosclerotic lesions. Using mouse and human S100β stem cells exposed to atherogenic stimuli in vitro, Tet-2 binding and histone modifications were assessed using quantitative single gene ChIP-PCR assays, genome-wide bulk ChIP-seq analysis, in-silico bulk ChIP-seq analysis qRT-PCR mRNA analysis. Single gene ChIP-PCR analysis revealed that S100β stem cells enrich for H3K27me3 at the SMC Myh11 locus but lose this mark and acquire the H3K4me2 mark with increased Tet-2 binding when undergoing myogenic differentiation in vitro, concomitant with increased expression of the Myh11 gene. Tet-2 siRNA knockdown attenuated the enrichment of H3K4me2 in response to atherogenic stimuli and reduced Myh11 gene expression. Adipogenic and osteogenic progeny, but not vasculogenic, also acquire this mark at the Myh11 locus without a parallel increase in Myh11 gene expression. Genome-wide bulk ChIP-seq analysis of H3K4me2 and H3K27me3 marks in human S100β cells revealed a similar profile while parallel in-silico bulk ChIP-seq analysis of published datasets for H3K4 methylation and H3K27me3 marks confirmed these data. Collectively, these data suggest the presence of the H3K4me2 mark at the Myh11 locus is not exclusive to de-differentiation of medial SMCs and may reflect the overall ‘commitment status’ of these cells and ultimately dictate their capacity to proceed and be fully determined and fated down specific lineages.