Adipose stem cells are sexually dimorphic cells with dual roles as preadipocytes and resident fibroblasts [bulk_adipocytes]

Cell identities are defined by intrinsic transcriptional networks and spatio-temporal environmental factors. Here, we explored multiple factors that contribute to the identity of adipose stem cells, including anatomic location, microvascular neighborhood, and sex. Our data suggest that adipose stem cells serve a dual role as adipocyte precursors and fibroblast-like cells that shape the adipose tissue's extracellular matrix in an organotypic manner. We further find that adipose stem cells display sexual dimorphism regarding genes involved in estrogen signaling, homebox transcription factor expression and the renin-angiotensin-aldosterone system. These differences could be attributed to sex hormone effects, developmental origin, or both. Finally, our data demonstrate that adipose stem cells are distinct from mural cells, and that the state of commitment to adipogenic differentiation is linked to their anatomic position in the microvascular niche. Our work supports the importance of sex and microvascular function in adipose tissue physiology. Overall design: To investigate the gene expression profile of mature adipocyte in male and female mice in pgWAT and iWAT depots, we isolated RNA from floating mature adipocytes (derived from collagenase treated tissue) and performed bulk RNA sequencing. There are five mouse RNAseq datasets connected to this study: (1) scRNAseq of stromal vascular fraction (SVF) of pgWAT, (2) bulk RNA-seq from FACS sorted Adipose stem cells (ASC) from pgWAT and iWAT, (3) bulk RNA-seq from isolated floating mature adipocytes from pgWAT and iWAT, (4) bulk RNA-seq from FACS sorted ASC from castrated/ovarectomized/male and female control mice, and (5) bulk RNA-seq from in vitro cultivated SVF cells from pgWAT and iWAT.