Project description:We used single cell sequencing to identify sex and diet-specific differences in visceral adipose progenitor cells (APCs) and to discover new APC markers and adipogenic modulators. We identified the major clusters that were previously identified by other studies both in humans and mice with the notable difference being in the mesothelial cluster, which was more abundant in humans than mice. We found that diet/obesity resulted in minor differences in the proportions of major clusters in humans but increased immune cells in male mice with high fat feeding. Most importantly, we identified BMPER as a novel marker of APCs both in humans and mice and revealed its pro-adipogenic property.
Project description:Background: Mature adipocytes are notoriously difficult to study ex vivo and alternative cell culture systems have therefore been developed. One of the most common models are human adipose progenitor cells (hAPCs). Unfortunately, these display replicative senescence after prolonged culture conditions, which limits their use in mechanistic studies. Methods: Herein, we knocked in human telomerase reverse transcriptase (TERT) into the AAVS1 locus of CD55+ hAPCs derived from abdominal subcutaneous adipose tissue and characterized the cells before and after differentiation into adipocytes. Results: Immortalized TERT-hAPCs retained proliferative and adipogenic capacities comparable to those of early-passage wild type hAPCs for >80 passages. In line with this, our integrative transcriptomic and proteomic analyses revealed that TERT-hAPCs displayed robust adipocyte expression profiles. This was confirmed by functional analyses of lipid turnover where TERT-hAPCs exhibited pronounced responses to insulin and pro-lipolytic stimuli such as isoprenaline, dibutyrul cAMP and tumor necrosis factor alpha. In addition, TERT-hAPCs could be readily cultured in both standard 2D and 3D-cultures and proteomic analyses revealed that the spheroid culture conditions improved adipogenesis. Conclusion: Through descriptive and functional studies, we demonstrate that immortalization of human CD55+ hAPCs is feasible and results in cells with stable proliferative and adipogenic capacities over multiple passages. As these cells are cryopreservable, they provide the additional advantage over primary cells of allowing repeated studies in both 2D and 3D model systems with the same genetic background.
Project description:We demonstrated the multipotent potential and change of cell identity of human adipose tissue-derived progenitors by subjecting cells to adipogenic, chondrogenic, and osteogenic induction. mRNA transcriptomic profiling results demonstrated the detection of lineage-specific markers.
Project description:To understand the specific transcriptomic changes in cells undergoing adipogenic differentiation, we performed single cell RNA-seq on two separate cultures: one corresponding to multipotent progenitors grown to confluency but not subjected to differentiation stimuli, and the second corresponding to progenitors exposed to adipogenic media for 3 days. Homogenous multipotent progenitors underwent two distinct fate trajectories rapidly upon induction of adipose differentiation, one toward the adipocyte fate, and the other towards a distinct, undifferentiated state.
Project description:Human adipose stem and progenitor cells (ASPCs) develop into heterogenous cultures of adipogenic and Structural Wnt-regulated Adipose Tissue resident (SWAT) cells upon induction of adipogenic differentiation. In vitro proliferating ASPC and differentiating adipocytes were collected from multiple timepoints to identify the trajectory of cells. Cells from two white depots (subcutaneous abdominal & visceral abdominal) and two brown depots (supraclavicular & perirenal) were used for the study. Progenitors from all 4 depots show similar differentiation trajectories during early differentiation.
Project description:human Multipotent Adipose-Derived Stem (hMADS) cells were subjected to adipogenic differentiation in vitro and microRNA expression was analyzed during differentiation.
Project description:human Multipotent Adipose-Derived Stem (hMADS) cells were subjected to adipogenic differentiation in vitro and microRNA expression was analyzed during differentiation. Total RNA was extracted at day 0 (AD0), day 3 (AD3) and day 8 (AD8) of differentiation, two biological replicates (1) and (2), and microRNA profiles were established with SOLiD sequencing.
Project description:In human dystrophies, the progressive muscle wasting is exacerbated by ectopic deposition of fat and fibrous tissue originating from fibro/adipogenic progenitors (FAPs). In degenerating muscles, the ability of these cells to adjuvate a successful healing is attenuated and FAPs aberrantly expand and differentiate into adipocytes and fibroblasts. Thus, arresting the fibroadipogenic fate of FAPs, without affecting their physiological role, represents a valuable therapeutic strategy for patients affected by muscle diseases. Here, using a panel of adipose progenitor cells including human-derived FAPs coupled with pharmacological perturbations and proteome profiling, we report that LY2090314 interferes with a genuine adipogenic program acting as WNT surrogate for the stabilization of a competent -catenin transcriptional complex. To predict the beneficial impact of LY2090314 in limiting ectopic deposition of fat in human muscles, we combined the Poly-Ethylene-Glycol-Fibrinogen biomimetic matrix with these progenitor cells to create a miniaturized 3D model of adipogenesis. Using this scalable system, we demonstrated that a two-digit nanomolar dose of this compound is effective to repress adipogenesis in a higher 3D scale, thus offering a concrete proof for the use of LY2090314 to limit FAP-derived fat infiltrates in dystrophic muscles.