Two and three-dimensional gene transfer from enzymatically degradable hydrogel scaffolds.
Two and three-dimensional gene transfer from enzymatically degradable hydrogel scaffolds.
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DOI：
10.1002/jemt.20840
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发表时间：
2010-09
影响因子：
2.5
通讯作者：
Segura, Tatiana
中科院分区：
文献类型：
--
作者：
Lei, Yuguo;Ng, Quinn K. T.;Segura, Tatiana
将间充质干细胞（MSC）接种在合成水凝胶支架内进行遗传修饰的能力将提供一种指导MSC分化的替代方法。在这份报告中，我们探讨了基因转移到骨髓间充质干细胞接种在顶部，或内基质金属蛋白酶（MMP）可降解的水凝胶，载有DNA/聚（乙烯亚胺）（PEI）聚合物。DNA/PEI复合物被封装在聚（乙二醇）（PEG）水凝胶内，通过迈克尔加成化学与MMP可降解肽交联。通过使用编码绿色荧光蛋白和荧光素酶的载体来可视化和定量基因转移。我们发现，基因转移到骨髓间充质干细胞是可能的细胞接种在二维和三维。二维和三维接种的细胞的荧光素酶表达量相似，即使三维接种的细胞数量显著更高，表明基因转移到二维接种的细胞比三维接种的细胞更有效。使用允许细胞浸润以递送DNA的水凝胶支架可以在体内产生持久的信号，这对于功能组织的再生是必不可少的。
The ability to genetically modify mesenchymal stem cells (MSCs) seeded inside synthetic hydrogel scaffolds would offer an alternative approach to guide MSC differentiation. In this report, we explored gene transfer to MSCs seeded on top, or inside matrix metalloproteinase (MMP) degradable hydrogels that were loaded with DNA/poly(ethylene imine) (PEI) polyplexes. DNA/PEI polyplexes were encapsulated inside poly(ethylene glycol) (PEG) hydrogels crosslinked with MMP degradable peptides via Michael Addition chemistry. Gene transfer was visualized and quantified through using a vector encoding for green fluorescent protein and luciferase. We found that gene transfer to MSCs was possible for cells seeded both in two and three dimensions. The amount of luciferase expression was similar for cells seeded in two and three dimensions even though the number of cells in three dimensions is significantly higher, indicating that gene transfer to cells seeded in two dimensions is more efficient than for cells seeded in three dimensions. The use of hydrogel scaffolds that allow cellular infiltration to deliver DNA may result in long lasting signals in vivo, which are essential for the regeneration of functional tissues.
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