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Efficient Gene Delivery to Mesenchymal Stem Cells by an Ethylenediamine‐Modified Polysaccharide from Mulberry Leaves
Author(s) -
Deng WenWen,
Cao Xia,
Wang Miao,
Yang Yan,
Su WeiYan,
Wei YaWei,
OuYang Zhen,
Yu JiangNan,
Xu XiMing
Publication year - 2012
Publication title -
small
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.785
H-Index - 236
eISSN - 1613-6829
pISSN - 1613-6810
DOI - 10.1002/smll.201101554
Subject(s) - lipofectamine , mesenchymal stem cell , transfection , gene delivery , polysaccharide , plasmid , in vitro , nucleic acid , ethyleneimine , microbiology and biotechnology , chemistry , genetic enhancement , cytotoxicity , biophysics , biochemistry , biology , gene , vector (molecular biology) , recombinant dna , organic chemistry , polymer
This study investigates the use of a natural polysaccharide isolated from mulberry leaves as a nonviral gene vector. Ethylenediamine is chemically grafted to the backbone of a polysaccharide from mulberry leaves (MPS) to acquire nucleic acid binding affinity. A particle‐size observation indicates that the cationic mulberry leaf polysaccharide (CMPS) can efficiently combine with plasmid transforming growth factor β1 (TGF‐β1) to form nanoscaled particles. In addition, the electrophoresis assay indicates a retarded plasmid migration when the CMPS/pTGF‐β1 weight ratio is increased to 30:1. The in vitro cell transfection experiment is performed based on bone marrow mesenchymal stem cells (MSCs) derived from rat femurs and tibias, and the findings reveal that the complex with a CMPS/pTGF‐β1 weight ratio of 50:1 exhibits the highest cell transfection effect, which is significantly higher than that of branched poly(ethyleneimine) (PEI) (25 kDa; p = 0.001, Student's t‐test) and slightly higher than Lipofectamine 2000. Moreover, the cytotoxicity assay also demonstrates that all of these tested complexes and the plasmid TGF‐β1 are nontoxic to mesenchymal stem cells (MSCs). The results of the living cell imaging confirm that more of the CMPS/plasmid TGF‐β1 nanoparticles can be taken up and at a faster rate by the MSCs than by the positive control Lipofectamine 2000; these data are consistent with the transfection efficiency data. Together, these results suggest that the CMPS/pTGF‐β1 nanoparticle can potentially be developed into a promising alternative for the transfer of therapeutic genes into cells.