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. 2010 Oct 15;26(2):778-83.
doi: 10.1016/j.bios.2010.06.025. Epub 2010 Jul 1.

Targeted nanoparticles enhanced flow electroporation of antisense oligonucleotides in leukemia cells

Shengnian Wang  1 , Xulang Zhang, Bo Yu, Robert J Lee, L James Lee
Affiliations

Targeted nanoparticles enhanced flow electroporation of antisense oligonucleotides in leukemia cells

Shengnian Wang et al. Biosens Bioelectron. .

Abstract

Liposome nanoparticles (LNs) with a targeting ligand were used in a semi-continuous flow electroporation (SFE) device to enhance in vitro delivery of exogenous oligonucleotides (ODN). Nanoparticles comprising transferrin-targeted lipoplex encapsulating ODN G3139 were mixed with K562 cells (a chronic myeloid leukemia cell line) and incubated for half an hour to accomplish nanoparticle binding. The mixture was then flowed through a SFE channel where electric pulses were given. Better ODN delivery efficiency was achieved with an increase of ∼24% to the case in combination of non-targeted LNs and SFE, and ∼60% to the case using targeted LNs alone, respectively. The MTS assay results confirmed cell viability greater than 75%.

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Figures

Figure 1
Figure 1
Schematic illustration on how Tf-LNs enhance flow-through electroporation through ligand-receptor binding on the cell membrane surface.
Figure 2
Figure 2
Schematic illustration of the Tf-LNs synthesis with an ethanol dilution approach.
Figure 3
Figure 3
Confocal microscopy images show the cellular binding of Tf-LNs G3139 on K562 cells. Cells were incubated with Tf-LNs encapsulating FITC-G3139 and fixed on poly-D-lysine coated cover glass slides. The scale bars represent 10 μm.
Figure 4
Figure 4
Phase contrast and fluorescence microscopic images of FITC-ODNs uptake by K562 cells, which were treated with (a) medium only (negative control); (b) free ODNs; nanotargeted FITC-ODNs encapsulated lipoplex nanoparticles, (c) non-Tf-LNs-G3139 alone, (d) non-Tf-LNs-G3139 + BE, and (e) non-Tf-LNs-G3139 + SFE; Tf-targeted FITC-ODNs encapsulated lipoplex nanoparticles, (f) Tf-LNs-G3139 alone, (g) Tf-LNs-G3139 + BE, and (h) Tf-LNs-G3139 + SFE. The FITC-ODNs concentration was set at 2 μM. BE: Bio-Rad Xcell Pulser Bulk Electorporation, 2 mm cuvette; SFE: semi-continuous flow electroporation. Electroporation conditions: unipolar square wave pulses with a pulse duration of 25 ms for both BE and SFE. The scale bars represent 100 μm.
Figure 5
Figure 5
Quantitatively analysis of the ODN delivery efficiency in K562 cells via flow cytometry (a) and the cell viability via MTS assay (b). The FITC-ODNs concentration was set at 2 μM. The error bars correspond to triplicate experiments made with independently produced batches of lipoplex nanoparticles. Electroporation conditions: unipolar square wave pulses with a pulse duration of 25 ms in both BE and SFE.
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