Label‐Free Sorting of Human Mesenchymal Stem Cells Using Insulating Dielectrophoresis

Human mesenchymal stem cells (hMSCs) are a multipotent yet heterogeneous cell population with immunosuppressive and regenerative properties, making them highly promising for stem cell therapies targeting metabolic diseases. However, the inherent heterogeneity of hMSCs presents challenges for producing consistent therapeutic outcomes, emphasizing the need to isolate functionally distinct subpopulations. In this study, we employed insulating dielectrophoresis (DEP) via a trap-and-release sorting strategy to generate and characterize subpopulations of adipose tissue (AT)-derived hMSCs. Voltage and frequency parameters were systematically tuned, revealing that higher voltages increased the percentage of trapped cells, while higher frequencies had less impact. Sorted cells underwent a 14-day adipogenic differentiation process, assessed by Oil Red O staining. Our results demonstrated that untrapped cell populations generated at lower voltage and frequency thresholds exhibited enhanced adipogenic differentiation compared to unsorted controls. These findings suggest that DEP can be leveraged to isolate progenitor cells within hMSC populations, enabling the production of homogeneous cell subsets with targeted functional potential. This work highlights the utility of insulating DEP for addressing hMSC heterogeneity and advancing the development of stem cell therapies.