title: Glucocorticoid Cell Priming Enhances Transfection Outcomes in Adult Human Mesenchymal Stem Cells.
abstract: Human mesenchymal stem cells (hMSCs) are one of the most widely researched stem cell types with broad applications from basic research to therapeutics, the majority of which require introduction of exogenous DNA. However, safety and scalability issues hinder viral delivery, while poor efficiency hinders nonviral gene delivery, particularly to hMSCs. Here, we present the use of a pharmacologic agent (glucocorticoid) to overcome barriers to hMSC DNA transfer to enhance transfection using three common nonviral vectors. Glucocorticoid priming significantly enhances transfection in hMSCs, demonstrated by a 3-fold increase in efficiency, 4-15-fold increase in transgene expression, and prolonged transgene expression when compared to transfection without glucocorticoids. These effects are dependent on glucocorticoid receptor binding and caused in part by maintenance of normal metabolic function and increased cellular (5-fold) and nuclear (6-10-fold) DNA uptake over hMSCs transfected without glucocorticoids. Results were consistent across five human donors and in cells up to passage five. Glucocorticoid cell priming is a simple and effective technique to significantly enhance nonviral transfection of hMSCs that should enhance their clinical use, accelerate new research, and decrease reliance on early passage cells.
authors.affiliation: Department of Biological Systems Engineering, University of Nebraska-Lincoln, Lincoln, Nebraska, USA.
authors.forename: Abby M
authors.initials: AM
authors.lastname: Kelly
authors.affiliation: Department of Biological Systems Engineering, University of Nebraska-Lincoln, Lincoln, Nebraska, USA.
authors.forename: Sarah A
authors.initials: SA
authors.lastname: Plautz
authors.affiliation: Department of Nutrition & Health Sciences, University of Nebraska-Lincoln, Lincoln, Nebraska, USA.
authors.forename: Janos
authors.initials: J
authors.lastname: Zempleni
authors.affiliation: Department of Biological Systems Engineering, University of Nebraska-Lincoln, Lincoln, Nebraska, USA. Electronic address: apannier2@unl.edu.
authors.forename: Angela K
authors.initials: AK
authors.lastname: Pannier
mesh_terms: D000328:Adult; D005260:Female; D015870:Gene Expression; D018014:Gene Transfer Techniques; D005938:Glucocorticoids / Q000494:pharmacology*; D006801:Humans; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects; D014162:Transfection / Q000379:methods*; D019076:Transgenes; D055815:Young Adult
chemical_list: D005938:Glucocorticoids