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. 2004 Feb 15;10(4):531-4.
doi: 10.3748/wjg.v10.i4.531.

Transfection efficiency of pORF lacZ plasmid lipopolyplex to hepatocytes and hepatoma cells

Xun Sun  1 , Hong-Wei Zhang, Zhi-Rong Zhang
Affiliations

Transfection efficiency of pORF lacZ plasmid lipopolyplex to hepatocytes and hepatoma cells

Xun Sun et al. World J Gastroenterol. .

Abstract

Aim: To develop a novel non-viral gene delivery system, which has a small particle size and a high transfection efficiency to hepatocyte and hepatoma cells.

Methods: Lipid-polycation-DNA lipopolyplex (LPD) was prepared by mixing plasmid DNA and polylysine. The resulted polyplex was incubated for 10 min at room temperature, following the addition of preformed cationic liposomes. The morphology of LPD was observed by transmission electron microscopy. The diameter and surface charge of LPD were measured by photon correlation spectroscopy (PCS). The nuclease protection ability of LPD was evaluated by agarose gel electrophoresis. Estimation of the transfection efficiency was performed by galactosidase assay in Chang cells and SMMC-7721 cells.

Results: LPD had a regular spherical surface. The average diameter and the zeta potential of LPD were 132.1 nm and 26.8 mV respectively. LPD could protect plasmid DNA from nuclease degradation after 2 hours incubation at 37 degrees while the naked DNA degraded rapidly. The average transfection efficiencies were 86.2+/-8.9% and 72.4+/-6.5% in Chang cells and SMMC-7721 cells respectively.

Conclusion: LPD has a rather small particle size and a high transfection activity. LPD may be a good non-viral vector for application in some gene delivery.

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Figures

Figure 1
Figure 1
Electronic transmission microscopy of LPD (× 35000).
Figure 2
Figure 2
Size distribution of LPD. The particle size was mea-sured by photon correlation spectroscopy (Malvern zetasizer 3000 HS). The average diameter of LPD is 132.1 nm.
Figure 3
Figure 3
Zeta potential of LPD. The zeta potential of LPD was 26.8 mV in 10 mM herps buffer (pH7.4).
Figure 4
Figure 4
Agarose gel electrophoresis of naked DNA and LPD subjected to DNase degradation. Lane 1: LPD incubated with DNase for 5 minutes. Lane 2: LPD incubated with DNase for 1 hour. Lane 3: LPD incubated with DNase for 2 hours. Lane 4: Naked DNA incubated with DNase for 5 minutes. Lane 5: Naked DNA incubated with DNase for 1 hour. Lane 6: Naked DNA incubated with DNase for 2 hours.
Figure 5
Figure 5
pORF LacZ lipopolyplex transfected Chang hepatocytes.
Figure 6
Figure 6
pORF LacZ lipopolyplex transfected SMMC-7721 hepatoma cells.
See this image and copyright information in PMC

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