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    Ppia and ywhaz constitute a stable pair of reference genes during electrical stimulation in mesenchymal stem cells

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    Publication date
    2022-01
    Author
    Steel, L.
    Ansell, David M.
    Amaya, E.
    Cartmell, S.H.
    Keyword
    Electrical stimulation
    Gene expression
    Housekeeping genes
    Mesenchymal stem cells
    Reference genes
    Reverse transcription-PCR
    Rights
    (c) 2022 The Authors. This is an Open Access article distributed under the Creative Commons CC-BY license (https://creativecommons.org/licenses/by/4.0/)
    Peer-Reviewed
    Yes
    Open Access status
    Gold
    
    Metadata
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    Abstract
    Mesenchymal stem cells (MSCs) are multipotent adult stem cells with great potential in regenerative medicine. One method for stimulating proliferation and differentiation of MSCs is via electrical stimulation (ES). A valuable approach for evaluating the response of MSCs to ES is to assess changes in gene expression, relative to one or more reference genes. In a survey of 25 publications that used ES on cells, 70% selected GAPDH as the reference gene. We conducted a study to assess the suitability of six potential reference genes on an immortalized human MSC line following direct current ES at seeding densities of 5000 and 10,000 cells/cm2 . We employed three methods to validate the most stable reference genes from qRT-PCR data. Our findings show that GAPDH and ACTB exhibit reduced stability when seeded at 5000 cell/cm2 . In contrast, we found that the most stable genes across both plating densities and stimulation regimes were PPIA and YWHAZ. Thus, in ES gene expression studies in MSCs, we support the use of PPIA and YWHAZ as an optimal reference gene pair, and discourage the use of ACTB and GAPDH at lower seeding densities. However, it is strongly recommended that similar verification studies are carried out based on cell type and different ES conditions.
    URI
    http://hdl.handle.net/10454/18729
    Version
    Published version
    Citation
    Steel L, Ansell DM, Amaya E et al (2022) Ppia and ywhaz constitute a stable pair of reference genes during electrical stimulation in mesenchymal stem cells. Applied Sciences (APPS). 12(1): 153.
    Link to publisher’s version
    https://doi.org/10.3390/app12010153
    Type
    Article
    Collections
    Life Sciences Publications

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