Department of Biomedical Engineering, University of California, Irvine, Irvine, CA 92697, USA; Department of Neurology, University of California, Irvine, Irvine, CA 92697, USA; Sue & Bill Gross Stem Cell Research Center, University of California, Irvine, Irvine, CA 92697, USA.
Department of Biomedical Engineering, University of California, Irvine, Irvine, CA 92697, USA; Department of Neurology, University of California, Irvine, Irvine, CA 92697, USA; Sue & Bill Gross Stem Cell Research Center, University of California, Irvine, Irvine, CA 92697, USA.
Methods. 2018 Jan 15;133:91-103. doi: 10.1016/j.ymeth.2017.08.016. Epub 2017 Aug 31.
Whole cell membrane capacitance is an electrophysiological property of the plasma membrane that serves as a biomarker for stem cell fate potential. Neural stem and progenitor cells (NSPCs) that differ in ability to form neurons or astrocytes are distinguished by membrane capacitance measured by dielectrophoresis (DEP). Differences in membrane capacitance are sufficient to enable the enrichment of neuron- or astrocyte-forming cells by DEP, showing the separation of stem cells on the basis of fate potential by membrane capacitance. NSPCs sorted by DEP need not be labeled and do not experience toxic effects from the sorting procedure. Other stem cell populations also display shifts in membrane capacitance as cells differentiate to a particular fate, clarifying the value of sorting a variety of stem cell types by capacitance. Here, we describe methods developed by our lab for separating NSPCs on the basis of capacitance using several types of DEP microfluidic devices, providing basic information on the sorting procedure as well as specific advantages and disadvantages of each device.
细胞膜电容是细胞膜的一种电生理特性,可作为干细胞命运潜能的生物标志物。通过介电泳 (DEP) 测量的细胞膜电容可区分在形成神经元或星形胶质细胞能力上存在差异的神经干细胞和祖细胞 (NSPC)。细胞膜电容的差异足以通过 DEP 富集形成神经元或星形胶质细胞的细胞,表明可以根据命运潜能通过细胞膜电容分离干细胞。通过 DEP 分选的 NSPC 无需标记,并且不会经历分选过程中的毒性作用。其他干细胞群体在分化为特定命运时也会出现细胞膜电容的变化,这阐明了通过电容对各种干细胞类型进行分选的价值。在这里,我们描述了我们实验室开发的几种基于电容的 DEP 微流控设备分离 NSPC 的方法,提供了有关分选程序的基本信息以及每种设备的具体优缺点。