Preparation of cationized polysaccharides as gene transfection carrier for bone marrow-derived mesenchymal stem cells.
The objective of this study is to prepare a non-viral carrier of gene transfection from various polysaccharides and evaluate the feasibility in gene expression for mesenchymal stem cells (MSCs). Various amounts of spermine were chemically introduced into pullulan, dextran and mannan with a molecular weight of around 40 000 or pullulan with different molecular weights to prepare cationized polysaccharides with different extents of spermine introduced (spermine-polysaccharide). Each cationized polysaccharide was complexed with a plasmid DNA at various ratios and in vitro gene transfection was investigated for rat bone marrow-derived MSCs. The level of gene expression depended on the type of cationized polysaccharide. The highest level was observed for the complex of spermine-pullulan and plasmid DNA. Additionally, the level also depended on the molecular weight of pullulan and the extent of spermine introduced to pullulan. Suppression of gene expression with chlorpromazine and methyl-beta-cyclodextrin of endocytosis inhibitors demonstrated that the cellular uptake of spermine-pullulan-plasmid DNA complexes was mediated by clathrin- and raft/caveolae-dependent endocytic pathways. The cationized pullulan is a promising non-viral carrier of plasmid DNA for MSCs.
Jo
Jun-ichiro
J
Department of Biomaterials, Institute for Frontier Medical Sciences, Kyoto University, 53 Kawara-cho Shogoin, Sakyo-ku, Kyoto 606-8507, Japan.
Okazaki
Arimichi
A
Nagane
Kentaro
K
Yamamoto
Masaya
M
Tabata
Yasuhiko
Y
D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D002470:Cell Survival / Q000187:drug effects; D004247:DNA / Q000235:genetics / Q000378:metabolism; D004705:Endocytosis / Q000187:drug effects; D005936:Glucans / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D008970:Molecular Weight; D010957:Plasmids / Q000235:genetics; D011134:Polysaccharides / Q000737:chemistry* / Q000378:metabolism* / Q000633:toxicity; D051381:Rats; D017208:Rats, Wistar; D013096:Spermine / Q000737:chemistry; D014162:Transfection / Q000379:methods*
D005936:Glucans; D011134:Polysaccharides; D013096:Spermine; C009109:pullulan; D004247:DNA