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阳离子化多糖的可生物降解明胶支架中的大鼠间充质基质细胞的基因转染。

A gene transfection for rat mesenchymal stromal cells in biodegradable gelatin scaffolds containing cationized polysaccharides.

机构信息

Department of Biomaterials, Institute for Frontier Medical Sciences, Kyoto University, Sakyo-ku, Kyoto, Japan.

出版信息

Biomaterials. 2011 Jan;32(3):919-25. doi: 10.1016/j.biomaterials.2010.09.056. Epub 2010 Oct 13.

Abstract

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.

摘要

本研究旨在设计能够对骨髓源性干细胞进行基因转染的三维支架,并评估其转染程度。制备了明胶和β-磷酸三钙(β-TCP)的三维支架。将引入亚精胺的普鲁兰(亚精胺-普鲁兰)作为基因转染的非病毒载体。将亚精胺-普鲁兰与荧光素酶质粒 DNA 混合,制备其聚离子复合物。用不同浓度的琥珀酰化明胶(suc-gel)处理支架,用不同的冷冻干燥和脱水热处理方法处理,然后将其置于复合物的水溶液中制备含有复合物的各种支架。当将干细胞接种到支架中以评估细胞的基因转染时,支架中包含复合物的制备方法决定了质粒 DNA 的转染水平。尽管释放曲线取决于支架的类型,但复合物随时间从支架中释放。支架中干细胞的基因转染顺序与释放的质粒 DNA 顺序非常一致。有可能是从支架中释放的复合物转染了细胞。

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