Functional Stratification of Adipose-Derived Stem Cells via Gradient Density Centrifugation: D-ASCs Drive Vascularization for Enhanced Fat Graft Survival.

Evidence from laboratory and animal studies provides groundwork for understanding mechanisms and potential before human studies continue.

• Level A · Stronger Clinical Evidence
• Level B · Emerging clinical evidence with positive signals
• Level C · Early human research exploring benefits
• Level D · Scientific groundwork from lab and animal studies
• Emerging · Emerging topic under active research

Abstract (original English)

To investigate the functional heterogeneity of adipose-derived stem cells (ASCs) in fat grafting and develop a density-based separation method to isolate distinct ASC subpopulations for improving graft survival. ASCs were isolated from human lipoaspirates and subjected to adipogenic differentiation. A novel Percoll density gradient centrifugation protocol (10-100%, 300 × g, 35 min) was developed to separate ASCs into upper-layer (U-ASCs) and lower-layer (D-ASCs) subpopulations. Their adipogenic and angiogenic potentials were assessed through Oil Red O staining and Matrigel tube formation assays. In vivo functionality was evaluated in nude mice (n = 84) receiving fat grafts supplemented with GFP-labeled U-ASCs, D-ASCs, or unsorted ASCs (n = 28/group), with graft retention, histology, and cellular fate analyzed over 12 weeks. Density separation yielded two functionally distinct populations: U-ASCs exhibited 3.2-fold greater lipid accumulation (p < 0.001), while D-ASCs showed 2.7-fold enhanced tube formation (p < 0.01). In vivo, D-ASCs grafts demonstrated 58.3 ± 6.7% retention at 12 weeks versus 32.1 ± 5.2% for U-ASCs (p < 0.001), with significantly reduced oil cysts (p < 0.05) and 2.4-fold higher vascular density (p < 0.001). Immunofluorescence revealed U-ASCs primarily differentiated into adipocytes (GFP+/Perilipin+), whereas D-ASCs integrated into vasculature (GFP+/CD31+). ASC

Evidence level

Evidence from laboratory and animal studies provides groundwork for understanding mechanisms and potential before human studies continue.

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