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A comparative evaluation of poly-l-lysine-palmitic acid and Lipofectamine ™ 2000 for plasmid delivery to bone marrow stromal cells

Paper ID Volume ID Publish Year Pages File Format Full-Text
10675 697 2007 12 PDF Available
Title
A comparative evaluation of poly-l-lysine-palmitic acid and Lipofectamine ™ 2000 for plasmid delivery to bone marrow stromal cells
Abstract

The current study compared the effectiveness of an amphiphilic biomaterial poly(l-lysine)-palmitic acid (PLL–PA), and the lipid-based transfection agent Lipofectamine™ 2000 for plasmid delivery to bone marrow stromal cells (BMSC). We investigated the utility of the carriers to deliver a plasmid containing enhanced green fluorescent protein (pEGFP) to BMSC in vitro. Confocal microscopy was used to investigate the intracellular trafficking of pEGFP/carrier complexes. pEGFP delivery and EGFP expression were assessed by flow cytometry. PLL–PA formed condensed structures with pEGFP and successfully delivered the plasmid into the nucleus within 5 h of incubation with the cells. PLL–PA delivered the pEGFP to ∼80% of the cells, achieving a maximum transfection efficiency of ∼22%. This was significantly higher than Lipofectamine™ 2000-mediated transfection, which was 11% under most optimal conditions. Dosing the BMSC two or three times during the 24 h period increased the transfection efficiency by 2–3 folds, without compromising cell viability. When chloroquine was employed as an ensomolytic agent, 100 μm of the drug increased the transfection efficiency while reducing cell viability, but lower concentrations (1–10 μm) were not beneficial for transfection. Combining PLL–PA with Lipofectamine™ 2000 created an additive effect, increasing the transfection efficiency of PLL–PA. Long-term evaluation of gene expression with pEGFP/PLL–PA yielded ∼17% transfection on day 1, which gradually decreased over a 12-day period. We conclude that PLL–PA is an effective biomaterial carrier and a promising candidate for non-viral gene delivery to BMSC.

Keywords
Gene transfer; Bone marrow; Mesenchymal stem cell; Polyamino acid; Poly(l-lysine); Palmitic acidAF546: Alexa Fluor 546 carboxylic acid succinimidyl ester; AdGFP: adenovirus expressing green fluorescent protein; BMSC: bone marrow stromal cells; EGFP: enhan
First Page Preview
A comparative evaluation of poly-l-lysine-palmitic acid and Lipofectamine ™ 2000 for plasmid delivery to bone marrow stromal cells
Publisher
Database: Elsevier - ScienceDirect
Journal: Biomaterials - Volume 28, Issue 31, November 2007, Pages 4693–4704
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering