RESEARCH ARTICLE


Electroporation Can Efficiently Transfect hESC-Derived Mesenchymal Stem Cells without Inducing Differentiation



Anthony J. Sprangers, Brian Freeman, Brenda M. Ogle*
University of Wisconsin- Madison, 2-114 Engineering Centers Building, 1550 Engineering Drive, Madison, WI 53706, USA.


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M. Ogle et al.

open-access license: This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International Public License (CC-BY 4.0), a copy of which is available at: https://creativecommons.org/licenses/by/4.0/legalcode. This license permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

* Address correspondence to this author at the University of Wisconsin- Madison, 2-114 Engineering Centers Building, 1550 Engineering Drive, Madison, WI 53706, USA. E-mail: ogle@wisc.edu


Abstract

Electroporation is a common method of gene transfer that has recently been used to efficiently transfect mesenchymal stem (stromal) cells (MSCs); however, the electrical stimulus has the potential to alter cell state. This study examines possible negative effects of electroporation of human embryonic stem cell (hESC)-derived MSCs including, loss of potency or induction of differentiation. Immunofluorescence and PCR were used to quantify protein and RNA expression of CD73 (an MSC marker) and markers of mature mesenchymal cell types following electroporation. The relative fraction of cells expressing CD73 protein was not altered in cells exposed to a 20 ms pulse at 1000 V or 1500 V compared to controls even after three passages, suggesting MSCs retain multipotency following electroporation. In addition, RNA expression of markers indicative of mature cells of bone, fat and cardiac muscle did not differ from unmanipulated controls soon after electroporation. Taken together, these results indicate electroporation under conditions favorable for MSC transfection does not significantly alter stem cell state.

Keywords: Electroporation, stem cells, mesenchymal stem cells, differentiation.