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In vivo endothelial commitment of adipose stem cells (ASCs) has scarcely been reported, and controversy remains on the contribution of ASCs to vascularization. We address the epigenetic commitment of ASCs to the endothelial lineage. We report a bisulfite sequencing analysis of CpG methylation in the promoters of two endothelial‐cell‐specific genes,  CD31  and  CD144 , in freshly isolated and in cultures of ASCs before and after induction of endothelial differentiation. In contrast to adipose tissue‐derived endothelial (CD31 + ) cells, freshly isolated ASCs display a heavily methylated  CD31  promoter and a mosaically methylated  CD144  promoter despite basal transcription of both genes. Methylation state of both promoters remains globally stable upon culture. Endothelial stimulation of ASCs in methylcellulose elicits phenotypic changes, marginal upregulation of  CD31 , and  CD144  expression and restrictive induction of a CD31 + CD144 +  immunophenotype. These events are accompanied by discrete changes in CpG methylation in  CD31  and  CD144  promoters; however, no global demethylation that marks CD31 +  cells and human umbilical vein endothelial cells occurs. Immunoselection of CD31 +  cells after endothelial stimulation reveals consistent demethylation of one CpG immediately 3′ of the transcription start site of the  CD31  promoter. Adipogenic or osteogenic differentiation maintains  CD31  and  CD144  methylation patterns of undifferentiated cells. Methylation profiles of  CD31  and  CD144  promoters suggest a limited commitment of ASCs to the endothelial lineage. This contrasts with the reported hypomethylation of adipogenic promoters, which reflects a propensity of ASCs toward adipogenic differentiation. Analysis of CpG methylation at lineage‐specific promoters provides a robust assessment of epigenetic commitment of stem cells to a specific lineage.  Disclosure of potential conflicts of interest is found at the end of this article.

stem cells

CpG Methylation Profiles of Endothelial Cell‐Specific Gene Promoter Regions in Adipose Tissue Stem Cells Suggest Limited Differentiation Potential Toward the Endothelial Cell Lineage