@article{0e89eee75cdf4ce5b4bb9124366dacbe,
title = "Ppia and ywhaz constitute a stable pair of reference genes during electrical stimulation in mesenchymal stem cells",
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.",
keywords = "Electrical stimulation, Gene expression, Housekeeping genes, Mesenchymal stem cells, Reference genes, Reverse transcription-PCR",
author = "Lynsey Steel and Ansell, {David M.} and Enrique Amaya and Cartmell, {Sarah H.}",
note = "Funding Information: This research was funded by the Engineering and Physical Sciences Research Council (EPSRC) and the Medical Research Council (MRC) Centre for Doctoral Training in Regenerative Medicine (EP/L014904/1). The authors wish to thank Stephen Richardson, Division of Cell Matrix Biology and Regenerative Medicine, The University of Manchester for gifting the Y201 cells, and his contribution to the design of experiments. Some of this work was conducted in the Henry Royce Institute for Advanced Materials, funded by the following EPSRC grants: EP/R00661X/1, EP/S019367/1, EP/P025021/1. The authors also wish to thank Katherine Bexley for assistance with the design of primers used in this study. Funding Information: Acknowledgments: The authors wish to thank Stephen Richardson, Division of Cell Matrix Biology and Regenerative Medicine, The University of Manchester for gifting the Y201 cells, and his contribution to the design of experiments. Some of this work was conducted in the Henry Royce Institute for Advanced Materials, funded by the following EPSRC grants: EP/R00661X/1, EP/S019367/1, EP/P025021/1. The authors also wish to thank Katherine Bexley for assistance with the design of primers used in this study. Funding Information: Funding: This research was funded by the Engineering and Physical Sciences Research Council (EPSRC) and the Medical Research Council (MRC) Centre for Doctoral Training in Regenerative Medicine (EP/L014904/1). Publisher Copyright: {\textcopyright} 2021 by the authors. Licensee MDPI, Basel, Switzerland.",
year = "2022",
month = jan,
day = "1",
doi = "10.3390/app12010153",
language = "English",
volume = "12",
journal = "Applied Sciences (Switzerland)",
issn = "2076-3417",
publisher = "MDPI",
number = "1",
}