A gene transfection for rat mesenchymal stromal cells in biodegradable gelatin scaffolds containing cationized polysaccharides.
The objective of this study was to design three-dimensional scaffolds of bone marrow-derived stem cells capable of their gene transfection and evaluate the transfection extent. Three-dimensional scaffolds of gelatin and β-tricalcium phosphate (β-TCP) were prepared. Spermine-introduced pullulan (spermine-pullulan) was prepared as the non-viral carrier of gene transfection. The spermine-pullulan was mixed with a luciferase plasmid DNA to prepare their polyion complex. The scaffolds were treated with succinylated gelatin (suc-gel) at different concentrations, treated in different methods of freeze-drying and dehydrothermal treatment, and placed in the aqueous of complexes to prepare various scaffolds containing the complex. When the stem cells were seeded into the scaffolds to evaluate the gene transfection of cells, the level of plasmid DNA transfection depended on the method of complex containing scaffolds preparation. The complexes were released with time from the scaffolds although the release profile depended on the type of scaffolds. The order of gene transfection for the stem cells in the scaffolds was in good accordance with that of plasmid DNA released. It is possible that cells were transfected with the complexes released from the scaffolds.
Kido
Yuichiro
Y
Department of Biomaterials, Institute for Frontier Medical Sciences, Kyoto University, Sakyo-ku, Kyoto, Japan.
Jo
Jun-ichiro
J
Tabata
Yasuhiko
Y
D000818:Animals; D002478:Cells, Cultured; D005780:Gelatin / Q000737:chemistry*; D005818:Genetic Engineering; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology*; D011134:Polysaccharides / Q000737:chemistry*; D051381:Rats; D017208:Rats, Wistar; D017154:Stromal Cells / Q000166:cytology* / Q000378:metabolism*; D054457:Tissue Scaffolds / Q000737:chemistry; D014162:Transfection / Q000379:methods*
D011134:Polysaccharides; D005780:Gelatin