Stem Cell Subpopulation Profiling as a Predictor of Human Health and Regenerative Capacity.
Michael W. Findlay, MBBS, PhD, FRACS, FACS, Michael Sorkin, MD, Sacha Khong, PhD, Robert C. Rennert, MD, Peter Than, MD, Michael Januszyk, MD, Christopher Davis, MBBCh, MRCS, Melanie Rodrigues, PhD, H. Peter Lorenz, MD, Homero Rivas, MD, John M. Morton, MD, Geoffrey C. Gurtner, MD.
Stanford University, Stanford, CA, USA.
Adipose-derived stem cells (ASC) are essential for tissue homeostasis and readily accessible for therapeutic purposes. Autologous therapy outcomes are variable despite refinements to harvest, processing and application techniques. We examined whether comorbidity-related depletion of ASC subpopulations could contribute to poor therapeutic responses and morbidity by examining ASCs at a single cell level.
The stromal vascular fraction (SVF) was isolated from humans (n=110) and mice (n=40) across a wide spectrum of age and comorbidities. Magnetic Assisted Cell Sorting (MACS) achieved lineage depletion before single cell sorting (CD45-CD31-CD34+) by Fluorescence-Activated Cell Sorting (FACS). Gene expression profiles were determined using 96 genes across a spectrum of stemness and related genes. Subpopulations were resolved by a fuzzy-C means cluster-based algorithm, then verified by surface marker profiling using density dependent (ViSNE) and independent (SPADE) analyses. Subpopulation profiles were correlated with 360 clinical variables over a median follow up of over 500 days.
Animal models of diabetes mellitus, obesity and aging demonstrated depletion of ASC subpopulations important for neovascularization- a novel cellular basis for the complication profiles of these states. These findings were confirmed in humans across a broad range of age (18-70years), BMI (24-78kg/m^2) and HBA1c (4.7-14%) with diabetes and obesity producing a significant 71-fold (p<0.0002) and 9-fold (p<0.0065) reduction in the CD26+CD55+ subpopulation respectively.
ASC subpopulation depletion is a novel mechanism for the variability seen in autologous therapies, correlates with the severity of comorbidities and raises questions as to the suitability of autologous ASC therapies in patients with these comorbidities.
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