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. 2013:2013:634379.
doi: 10.1155/2013/634379. Epub 2013 Aug 21.

Aldehyded Dextran and ε -Poly(L-lysine) Hydrogel as Nonviral Gene Carrier

Yumiko Togo  1 , Katsu Takahashi, Kazuyuki Saito, Honoka Kiso, Boyen Huang, Hiroko Tsukamoto, Suong-Hyu Hyon, Kazuhisa Bessho
Affiliations

Aldehyded Dextran and ε -Poly(L-lysine) Hydrogel as Nonviral Gene Carrier

Yumiko Togo et al. Stem Cells Int. 2013.

Abstract

Background. The expression term of the gene transfected in cells needs to belong enough inorder to make a gene therapy clinically effective. The controlled release of the transfected gene can be utilized. The new biodegradable hydrogel material created by 20 w/w% aldehyded dextran and 10 w/w% ε -poly(L-lysine) (ald-dex/PLL) was developed. We examined whether it could be as a nonviral carrier of the gene transfer. Methods. A plasmid (Lac-Z) was mixed with ald-dex/PLL. An in vitro study was performed to assess the expression of Lac-Z with X-gal stain after gene transfer into the cultured 293 cells and bone marrow cells. As a control group, PLL was used as a cationic polymer. Results. We confirmed that the transfection efficiency of the ald-dex/PLL had a higher transfection efficiency than PLL in 293 cells (plasmid of 2 μ g: ald-dex/PLL 1.1%, PLL 0.23%, plasmid of 16 μ g: ald-dex/PLL 1.23%, PLL 0.48%). In bone marrow cells, we confirmed the expression of Lac-Z by changing the quantity of aldehyded dextran. In the groups using ald-dextran of the quantity of 1/4 and 1/12 of PLL, their transfection efficiency was 0.43% and 0.41%, respectively. Conclusions. This study suggested a potential of using ald-dex/PLL as a non-carrier for gene transfer.

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Figures

Figure 1
Figure 1
The chemical structure and cross-linking of the new biodegradable hydrogel composed of aldehyded dextran/PLL and plasmid DNA. After the mixture of the solution, gel formation proceeds on the basis of Schiff base formation. Plasmid DNA interacts with PLL by electrostatic interactions between positively charged amine of the PLL and negatively charged phosphate groups of the plasmid DNA.
Figure 2
Figure 2
Gene transfection into 293 cells. (a), (b), and (c): results of gene transfection in plasmid of 2 μg. (d), (e), and (f): results of gene transfection in plasmid of 16 μg. (a) and (d) were carried out by only plasmid, and (b) and (e) were carried out by using PLL as a gene carrier, for the purpose of controls. (c) and (f) were carried out by using ald-dex/PLL as a gene carrier.
Figure 3
Figure 3
Gene transfection into MSCs. (a) Transfection with ald-dex (1/1 of PLL) and PLL. (b) Transfection with ald-dex (1/4 of PLL) and PLL. (c) Transfection with ald-dex (1/12 of PLL) and PLL.
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