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Detection of transfected cells by confocal microscopy
€
ed. F. Vogtle, G. Richardt and N. Werner, Wiley-VCH, Weinheim,
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In order to observe the transfected osteoblasts after magneto-
fection, cells were examined by a Biorad MC1025 Confocal Laser
Scanning Microscope. The fluorescence of EGFP was excited at
488 nm and the emitted fluorescence was measured at 530/30 nm.
Cell viability was determined by addition of propidium iodide
(PI; 0.005% in PBS, Sigma-Aldrich Corporation, St. Louis, MO,
USA) to stain the DNA of dead cells.
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Conclusions
In this article we present a novel nonviral vector designed for
DNA magnetofection based on PPI dendrimers and magnetic
nanoparticles. The dendrimers are attached to the nanoparticles
in one synthetic step, without any additional modification of the
iron oxide surface. The nanosystem is chemically designed with
a covalent bond between both components, thus ensuring the
vector stability from the extracellular environment up to the cell
interior. This strategy provides an important advantage, espe-
cially as a genetic engineering tool since DNA can be delivered in
a short time while keeping the structure integrity. The trans-
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generation to protect and transport the new gene into the
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Acknowledgements
This work was supported by the Spanish CICYT through the
project MAT2008-00736 and by the CAM through project
S2009/MAT-1472. We thank C. Aroca and M. Maicas (ISOM,
ꢀ
Universidad Politecnica de Madrid) for their technical assistance
in the magnetic measurements.
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