Analogous to the classical Th1, Th2, and Th17 helper T cell subsets, NKT1, NKT2, and NKT17 cells have been described with stereotypical cytokine production profiles, transcription factor expression, and tissue localization. Many reports have described iNKT cells in the context of proinflammatory immune responses where they respond to danger signals and microbial lipid antigens to mediate host defense. In contrast, we recently identified a distinct role of iNKT cells in adipose tissues of mice and humans where they display a unique regulatory phenotype. Adipose iNKT cells produce high levels of IL-2 and IL-10, which drive the expansion of Tregs and M2 macrophages, respectively, and have a unique transcriptional profile that sets them apart from iNKT cells in other organs. How these iNKT cells arise, however, is incompletely understood. We performed coculture of splenic and thymic iNKT cells with adipose tissue, and found that they rapidly adopted many of the transcription factor, surface marker, and cytokine production profiles characteristic of adipose iNKT cells, including an upregulation of E4BP4 and a downregulation of PLZF. We then performed in vitro and in vivo analyses to identify the cellular and molecular inducers of regulatory iNKT cells in adipose tissue. Our work highlights the importance of the adipose microenvironment in contributing to the unique phenotype of adipose iNKT cells.