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Efficient Nonviral Transfection of Human Bone Marrow Mesenchymal Stromal Cells Shown Using Placental Growth Factor Overexpression
Joint Authors
Cheung, Winston Y.
Hovey, Owen
Gobin, Jonathan M.
Muradia, Gauri
Mehic, Jelica
Westwood, Carole
Lavoie, Jessie R.
Source
Issue
Vol. 2018, Issue 2018 (31 Dec. 2018), pp.1-10, 10 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2018-12-24
Country of Publication
Egypt
No. of Pages
10
Abstract EN
Background.
Human mesenchymal stromal/stem cells (hMSCs) hold great therapeutic potential due to their immunomodulatory and tissue regenerative properties.
Enhancement of biological features of hMSCs by transfection has become a focus of investigation for cell- and gene-based therapies.
However, many of the current transient transfection methods result in either low transfection efficiency or high cytotoxicity.
Methods.
In order to find a transfection method that would address the current issues of low transfection efficiency and high cytotoxicity, 6 commercially available cationic lipid and polymer reagents were tested on human bone marrow-derived MSCs (hBM-MSCs) using GFP as a reporter gene.
One transfection method using TransIT-2020 was selected and tested with an emphasis on cell quality (viability, identity, and yield), as well as efficacy with a human placental growth factor (PlGF) plasmid.
Results.
TransIT-2020 yielded the highest fluorescence signal per cell out of the methods that did not decrease cell recovery.
Transfecting GFP to 5 hBM-MSC donors using TransIT-2020 yielded 24–36% GFP-expressing cells with a viability of 85–96%.
hBM-MSC identity was unaffected as CD90, CD105, and CD73 markers were retained (>95%+) after transfection.
When this method was applied to PlGF expression, there was up to a 220-fold increase in secretion.
Both growth and secretion of PlGF in overexpressing hBM-MSC were sustained over 7 days, confirming the sustainability and applicability of the TransIT-2020 transfection system.
Discussion.
We report a simple and efficient method for transient transfection that has not been reported for hBM-MSCs, encompassing high levels of plasmid expression without significant changes to fundamental hBM-MSC characteristics.
American Psychological Association (APA)
Cheung, Winston Y.& Hovey, Owen& Gobin, Jonathan M.& Muradia, Gauri& Mehic, Jelica& Westwood, Carole…[et al.]. 2018. Efficient Nonviral Transfection of Human Bone Marrow Mesenchymal Stromal Cells Shown Using Placental Growth Factor Overexpression. Stem Cells International،Vol. 2018, no. 2018, pp.1-10.
https://search.emarefa.net/detail/BIM-1213139
Modern Language Association (MLA)
Cheung, Winston Y.…[et al.]. Efficient Nonviral Transfection of Human Bone Marrow Mesenchymal Stromal Cells Shown Using Placental Growth Factor Overexpression. Stem Cells International No. 2018 (2018), pp.1-10.
https://search.emarefa.net/detail/BIM-1213139
American Medical Association (AMA)
Cheung, Winston Y.& Hovey, Owen& Gobin, Jonathan M.& Muradia, Gauri& Mehic, Jelica& Westwood, Carole…[et al.]. Efficient Nonviral Transfection of Human Bone Marrow Mesenchymal Stromal Cells Shown Using Placental Growth Factor Overexpression. Stem Cells International. 2018. Vol. 2018, no. 2018, pp.1-10.
https://search.emarefa.net/detail/BIM-1213139
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1213139