Department of Chemical and Biomolecular Engineering, University of California, Irvine, Irvine, CA, 92697-2580, USA; Department of Neurology, University of California, Irvine, Irvine, CA, 92697-6750, USA; Sue & Bill Gross Stem Cell Research Center, University of California, Irvine, CA, 92697-1705, USA.
Department of Biomedical Engineering, University of California, Irvine, Irvine, CA, 92697-2627, USA; Department of Neurology, University of California, Irvine, Irvine, CA, 92697-6750, USA; Sue & Bill Gross Stem Cell Research Center, University of California, Irvine, CA, 92697-1705, USA.
Biosens Bioelectron. 2020 Mar 15;152:111982. doi: 10.1016/j.bios.2019.111982. Epub 2019 Dec 28.
Human neural stem and progenitor cells (hNSPCs) have therapeutic potential to treat neural diseases and injuries since they provide neuroprotection and differentiate into astrocytes, neurons, and oligodendrocytes. However, cultures of hNSPCs are heterogeneous, containing cells linked to distinct differentiated cell fates. HNSPCs that differentiate into astrocytes are of interest for specific neurological diseases, creating a need for approaches that can detect and isolate these cells. Astrocyte-biased hNSPCs differ from other cell types in electrophysiological properties, namely membrane capacitance, and we hypothesized that this could be used to enrich these cells using dielectrophoresis (DEP). We implemented a two-step DEP sorting scheme, consisting of analysis to define the optimal sorting frequency followed by separation of cells at that frequency, to test whether astrocyte-biased cells could be separated from the other cell types present in hNSPC cultures. We developed a novel device that increased sorting reproducibility and provided both enriched and depleted cell populations in a single sort. Astrocyte-biased cells were successfully enriched from hNSPC cultures by DEP sorting, making this the first study to use electrophysiological properties for label-free enrichment of human astrocyte-biased cells. Enriched astrocyte-biased human cells enable future experiments to determine the specific properties of these important cells and test their therapeutic efficacy in animal models of neurological diseases.
人神经干细胞和祖细胞（hNSPCs）具有治疗神经疾病和损伤的潜力，因为它们提供神经保护并分化为星形胶质细胞、神经元和少突胶质细胞。然而，hNSPC 培养物是异质的，包含与不同分化细胞命运相关的细胞。分化为星形胶质细胞的 hNSPCs 对特定的神经疾病具有重要意义，因此需要能够检测和分离这些细胞的方法。与其他细胞类型相比，星形胶质细胞偏向的 hNSPCs 在电生理特性方面存在差异，即膜电容，我们假设这可以用于使用电（DEP）来富集这些细胞。我们实施了两步 DEP 分选方案，包括分析以确定最佳分选频率，然后在该频率下分离细胞，以测试星形胶质细胞偏向的细胞是否可以与 hNSPC 培养物中存在的其他细胞类型分离。我们开发了一种新型设备，提高了分选的重现性，并在单次分选中提供了富集和耗尽的细胞群体。通过 DEP 分选成功地从 hNSPC 培养物中富集了星形胶质细胞偏向的细胞，这是第一项使用电生理特性进行无标记人星形胶质细胞偏向细胞富集的研究。富集的星形胶质细胞偏向人细胞使未来的实验能够确定这些重要细胞的特定特性，并在神经疾病的动物模型中测试它们的治疗效果。
