Plastic Surgery Research Council
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IDENTIFICATION OF A FUNCTIONALLY DISTINCT SUBPOPULATION OF HUMAN ADIPOSE DERIVED STEM CELLS USING SINGLE CELL TRANSCRIPTIONAL ANALYSIS
Presenter: Michael Sorkin, MD
Co-Authors: Rennert R; Januszyk M; Chung MT; Longaker MT; Gurtner GC
Stanford University

Introduction: Human adipose-derived stem cells (hASCs) are clinically appealing due to their ease of harvest and multipotency. However, a more detailed understanding of their heterogeneity is crucial for the development of targeted clinical applications. Here, we utilize single cell microfluidic technology in conjunction with FACS to identify and prospectively isolate a subpopulation of hASCs with enhanced functional properties.

Methods: Primary hASCs were isolated from lipoaspirates, and microfluidic-based single cell transcriptional analysis was employed to simultaneously characterize the expression of 96 stemness, differentiation and surface antigen genes. A novel bioinformatic clustering analysis was used to identify hASC subpopulations based on transcriptional signatures. A selected subpopulation was prospectively isolated using FACS for assessment of enhanced in vitro function.

Results: Significant transcriptional heterogeneity was observed within hASCs, which were clustered based on expression profiles. One subpopulation (~10% of cells) displayed the coordinated over-expression of multiple stemness-associated and functional genes, and was correlated with the expression of selected cell surface markers. FACS and repeat transcriptional analyses confirmed the reproducibility and surface marker fidelity of this subpopulation across multiple patients. Prospective isolation resulted in the prolonged retention of progenitor associated surface antigens, as well as an enhanced proliferative capacity and clonogenicity when compared to negatively selected and parent populations. Additionally, matrigel co-culture of the identified hASC subpopulation with HUVEC cells resulted in a significant increase in HUVEC tubule formation.

Conclusion: Functionally distinct hASC subpopulations can be transcriptionally identified and linked to surface marker expression for prospective isolation and applications. Here, we isolated a subpopulation with superior functional properties. Further characterization of this hASC subpopulation, as well as other stem cell pools, may lead to more targeted cell-based therapies.


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