Label-free enrichment of fate-biased human neural stem and progenitor cells

Tayloria N G Adams; Alan Y L Jiang; Nicolo S Mendoza; Clarissa C Ro; Do-Hyun Lee; Abraham P Lee; Lisa A Flanagan

doi:10.1016/j.bios.2019.111982

Label-free enrichment of fate-biased human neural stem and progenitor cells

Biosens Bioelectron. 2020 Mar 15:152:111982. doi: 10.1016/j.bios.2019.111982. Epub 2019 Dec 28.

Authors

Tayloria N G Adams¹, Alan Y L Jiang², Nicolo S Mendoza³, Clarissa C Ro³, Do-Hyun Lee⁴, Abraham P Lee⁴, Lisa A Flanagan⁵

Affiliations

¹ 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. Electronic address: tayloria@uci.edu.
² 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.
³ 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.
⁴ Department of Biomedical Engineering, University of California, Irvine, Irvine, CA, 92697-2627, USA.
⁵ 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; Department of Anatomy & Neurobiology, University of California, Irvine, Irvine, CA, 92697-4291, USA. Electronic address: lisa.flanagan@uci.edu.

Abstract

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.

Keywords: Astrocyte; Cell sorting; Dielectrophoresis; Membrane capacitance; Microfluidic; Neural stem and progenitor cell.

MeSH terms

Astrocytes / cytology
Biosensing Techniques / instrumentation
Cell Line
Cell Separation / instrumentation*
Electric Capacitance
Equipment Design
Humans
Lab-On-A-Chip Devices*
Neural Stem Cells / cytology*
Neurons / cytology
Oligodendroglia / cytology

Grants and funding

UL1 TR000153/TR/NCATS NIH HHS/United States