Adipose-derived Stem cll and Stronal Vascular Fraction response to inflammation sc RNA seq

The therapeutic potential of mesenchymal stem/stromal cells (MSCs) relies on their ability to modulate immune responses in disease and wound contexts. In this study, we present an analytical pipeline using established algorithms to better characterize the response of adipose-derived stem/stromal cells (ASCs) to in vitro inflammatory stimuli, which in turn has therapeutic benefit in the context of MSC cell therapies. By integrating single-cell RNA sequencing and bulk proteomics data of ASCs derived from the stromal vascular fraction of adipose tissue, we present a detailed characterization of the ASC response to inflammation and additionally introduce a method of relating phenotypes observed in exogenously stimulated culture conditions to subpopulations of in vivo cells. Our pipeline involves multiple benchmarks to establish concordance between single-cell and bulk assays to more comprehensively characterize the ASC response to inflammation. This study is the introduction of a practical and efficient approach to identify functionally homogeneous and therapeutically relevant cell populations from heterogeneous mixtures using computational strategies and provides deeper insight into the ASC response to inflammation. Overall design: Stromal vascular fraction derived from human adipose tissue, as welll as passaged control (ctrl) and cytokine-stimulated (Stim) ASCs from one of the SVF donors. Stim cells were cultured in IFNg and TNFa fro 48 hours. Samples were all run on the 10x genomics single cell RNA sequencing platform.