Improved transfection in human mesenchymal stem cells: effective intracellular release of pDNA by magnetic polyplexes.
OBJECTIVE
Magnetically guided transfection has been shown as a promising approach for the genetic modification of cells. We observed that polyethylenimine (PEI)-condensed pDNA, combined with magnetic nanoparticles (MNPs) via biotin-streptavidin interactions could provide higher transfection efficiency than pDNA/PEI alone, even without the application of a magnetic force. Therefore, we intended to investigate the beneficial properties of MNP-based transfection.


METHODS
We performed three-color fluorescent labeling of magnetic transfection complexes and traced them inside human mesenchymal stem cells over time using confocal microscopy in order to study pDNA release kinetics by colocalization studies.


RESULTS
We demonstrated that MNP-combined pDNA/PEI complexes provide more rapid and efficient release of pDNA than pDNA/PEI alone, which could be explained by the retention of PEI on the surface of the MNPs due to strong biotin-streptavidin interactions.


CONCLUSIONS
The process of pDNA liberation may significantly influence the efficiency of the transfection vector. Therefore, it should be carefully considered when creating novel gene delivery agents.
Reference & Translation Center for Cardiac Stem Cell Therapy, Department of Cardiac Surgery, University of Rostock, Schillingallee 35, 18057 Rostock, Germany.
Evgenya
E
Delyagina
Anna
A
Schade
Dorothee
D
Scharfenberg
Anna
A
Skorska
Cornelia
C
Lux
Wenzhong
W
Li
Gustav
G
Steinhoff
D001710:Biotin / Q000737:chemistry; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000378:metabolism*; D010957:Plasmids / Q000737:chemistry* / Q000235:genetics; D011094:Polyethyleneimine / Q000737:chemistry; D019809:Streptavidin / Q000737:chemistry; D014162:Transfection / Q000379:methods*
D001710:Biotin; D011094:Polyethyleneimine; D019809:Streptavidin
colocalization study; gene delivery; magnetic nanoparticle; mesenchymal stem cell; polyethylenimine