Isolation of mature adipocytes from in vitro differentiated human adipose stem cells
Obesity and associated metabolic outcomes define the metabolic syndrome. Interestingly, an under-appreciated fact is that body fat distribution, rather than total body fat amount, is a key determinant of metabolic disease. Indeed, in contrast to upper-body obesity, lower-body fat accumulation inversely correlates with metabolic risks. Understanding processes regulating upper- vs. lower-body fat expansion is paramount to predict (and prevent) these risks. We combine functional, proteomics, transcriptomics and epigenomics analyses to identify chromatin-associated mechanisms of adipose depot-specific fat expansion. Here, we analyze by RNA-seq the transcriptome of adipocytes differentiated in vitro from human gluteal (lower-body) and abdominal subcutaneous (upper-body) depots-derived adipose stem cells. We aim to identify adipose depot-specific and temporal differences in the up- or down-regulation of gene expression, and relate these differences to changes in chromatin states. Overall design: Analysis of the transcriptome by RNA-seq in adipose tissue-derived stem cells (ASCs) induced to differentiate in vitro into the adipocyte lineage. ASCs were collected at the proliferating stage, day 0 (confluent, cell cycle-arrested stage, time of differentiation induction), and on days 1, 3, 9 and 15 of differentiation. ASCs were isolated from two donors (1 from gluteofemoral subcutaneous adipose tisse, 1 from abdominal subcutaneous adipose tissue), cultured and subjected to three independent differentiation experiments for each donor. Cells were collected at indicated time points and processed for RNA-seq analysis.  Code for data processing and analysis are available at https://github.com/sarahhp/nucleolus_paper.