Enrichment of differentiated hNT neurons and subsequent analysis using flow-cytometry and xCELLigence sensing

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dc.contributor.author MacDonald, C en
dc.contributor.author Unsworth, Charles en
dc.contributor.author Graham, Euan en
dc.date.accessioned 2018-10-10T00:56:57Z en
dc.date.issued 2014-04-30 en
dc.identifier.issn 0165-0270 en
dc.identifier.uri http://hdl.handle.net/2292/40236 en
dc.description.abstract Background: Human neurons (hNT neurons), obtained from the NTera2/D1 precursor cell line, are highly valued by many neuroscientists as isolation of adult human primary neuronal cells continues to elude us. hNT neurons are generated by differentiation of the NT2 precursors for a period of 4 weeks followed by 2 weeks of mitotic inhibition. This yields a heterogeneous population of neuronal phenotypes and underlying astrocyte precursors, the latter of which are very difficult to visualise using standard light microscopy. Such a mixed culture is acceptable for some applications (e.g. measurement of synaptic plasticity), whereas others (e.g. proteomics or transcriptomics) require almost pure cultures of hNT neurons. New method: Here we describe a simple method for obtaining highly enriched cultures of hNT neurons following the first neuronal harvest and detail several additional methods, namely flow-cytometry and xCELLigence© biosensor technology, to rapidly and reliably determine the purity and viability of the cultures. Comparison with existing methods: This method of enrichment for the neurons is novel and advances the end user applications of the cells. Results: In addition, we apply the enrichment method to conduct analysis of cell-surface markers using flow-cytometry on the enriched neuronal cells. Furthermore, we apply this method to generate enriched neuronal cells on which we conduct analysis of cell-surface markers using flow-cytometry. Conclusions: Collectively, this paper describes several new advances, which will create opportunities when using these cells and similar preparations, and provides the protocol for analysis of these cells using flow-cytometry and biosensor technology. © 2014 Elsevier B.V. en
dc.relation.ispartofseries Journal of Neuroscience Methods en
dc.rights Items in ResearchSpace are protected by copyright, with all rights reserved, unless otherwise indicated. Previously published items are made available in accordance with the copyright policy of the publisher. en
dc.rights.uri https://researchspace.auckland.ac.nz/docs/uoa-docs/rights.htm en
dc.title Enrichment of differentiated hNT neurons and subsequent analysis using flow-cytometry and xCELLigence sensing en
dc.type Journal Article en
dc.identifier.doi 10.1016/j.jneumeth.2014.02.004 en
pubs.begin-page 47 en
pubs.volume 227 en
dc.rights.holder Copyright: The author en
pubs.end-page 56 en
pubs.publication-status Published en
dc.rights.accessrights http://purl.org/eprint/accessRights/RestrictedAccess en
pubs.subtype Journal Article en
pubs.elements-id 434810 en
pubs.org-id Engineering en
pubs.org-id Engineering Science en
pubs.org-id Medical and Health Sciences en
pubs.org-id Medical Sciences en
pubs.org-id Molecular Medicine en
dc.identifier.eissn 1872-678X en

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