Neural stem and progenitor cells (NSPCs) are essential for brain development and represent a promising source for regenerative therapies. However, intrinsic heterogeneity within NSPC populations complicates the isolation of lineage-biased progenitors, particularly in the absence of reliable surface markers. Here, we apply dielectrophoresis (DEP), a label-free method that separates viable cells based on intrinsic electrophysiological properties, to enrich neuronal subtype–biased progenitors from embryonic mouse forebrain.
Using a custom multi-outlet microfluidic platform, NSPCs isolated from embryonic day 15.5 cortex and ganglionic eminence were sorted according to their electrophysiological profiles. Neuron-biased progenitors were distributed across multiple outlets, revealing heterogeneity within neuronal-primed populations. Post-sort immunocytochemistry and gene expression analyses confirmed enrichment of βIII-tubulin–positive cells and an increased proportion of GABA-expressing neurons following differentiation in specific fractions with distinct electrophysiological characteristics.
Neuronal markers were detected across all fractions, supporting the presence of diverse neuronally biased subpopulations. These findings demonstrate that intrinsic electrophysiological properties can serve as functional indicators of lineage bias and establish DEP as a scalable, label-free strategy to resolve and enrich heterogeneous neuronal progenitor subtypes for developmental studies and therapeutic applications. Research reported in this publication was supported in part by NSF CAREER Award IOS-1254060 (to L.A.F.), the National Institute of Neurological Disorders and Stroke of the National Institutes of Health under Award Number T32NS082174 (J.S.M.), and the Sue and Bill Gross Hall Research Center.
The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. The authors declare no conflict of interest. Department of Anatomy & Neurobiology, University of California, Irvine, Irvine, CA, 92697-4291, USA Jazmine S.
Adams Sue & Bill Gross Stem Cell Research Center, University of California, Irvine, Irvine, CA, 92697-1705, USA Jazmine S. Adams Department of Neurology, University of California, Irvine, Irvine, CA, 92697-6750, USA Department of Biomedical Engineering, University of California, Irvine, Irvine, CA, 92697-2627, USA Department of Chemical and Biomolecular Engineering, University of California, Irvine, Irvine, CA, 92697-2580, USA The authors declare no competing interests. Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
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