title: Environmental parameters influence non-viral transfection of human mesenchymal stem cells for tissue engineering applications.
abstract: Non-viral transfection is a promising technique that could be used to increase the therapeutic potential of stem cells. The purpose of this study was to explore practical culture parameters of relevance in potential human mesenchymal stem cell (hMSC) clinical and tissue engineering applications, including type of polycationic transfection reagent, N/P ratio and dose of polycation/pDNA polyplexes, cell passage number, cell density and cell proliferation. The non-viral transfection efficiency was significantly influenced by N/P ratio, polyplex dose, cell density and cell passage number. hMSC culture conditions that inhibited cell division also decreased transfection efficiency, suggesting that strategies to promote hMSC proliferation may be useful to enhance transfection efficiency in future tissue engineering studies. Non-viral transfection treatments influenced hMSC phenotype, including the expression level of the hMSC marker CD105 and the ability of hMSCs to differentiate down the osteogenic and adipogenic lineages. The parameters found here to promote hMSC transfection efficiency, minimize toxicity and influence hMSC phenotype may be instructive in future non-viral transfection studies and tissue engineering applications.
authors.affiliation: Department of Biomedical Engineering, University of Wisconsin-Madison, Madison, WI, USA.
authors.forename: William J
authors.initials: WJ
authors.lastname: King
authors.forename: Nicholas A
authors.initials: NA
authors.lastname: Kouris
authors.forename: Siyoung
authors.initials: S
authors.lastname: Choi
authors.forename: Brenda M
authors.initials: BM
authors.lastname: Ogle
authors.forename: William L
authors.initials: WL
authors.lastname: Murphy
mesh_terms: D000818:Animals; D015703:Antigens, CD / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D003470:Culture Media / Q000494:pharmacology*; D004247:DNA / Q000378:metabolism; D000071063:Endoglin; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D041681:NIH 3T3 Cells; D010957:Plasmids / Q000378:metabolism; D011094:Polyethyleneimine / Q000737:chemistry; D011956:Receptors, Cell Surface / Q000378:metabolism; D023822:Tissue Engineering / Q000379:methods*; D014162:Transfection / Q000379:methods*; D014780:Viruses / Q000187:drug effects / Q000378:metabolism
chemical_list: D015703:Antigens, CD; D003470:Culture Media; C064206:ENG protein, human; D000071063:Endoglin; D011956:Receptors, Cell Surface; D011094:Polyethyleneimine; D004247:DNA