Effective Gene Delivery to Mesenchymal Stem Cells Based on the Reverse Transfection and Three-Dimensional Cell Culture System
Effective Gene Delivery to Mesenchymal Stem Cells Based on the Reverse Transfection and Three-Dimensional Cell Culture System
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基于反转染和三维细胞培养系统的间充质干细胞有效基因递送
DOI：
10.1007/s11095-011-0390-0
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发表时间：
2011-02
影响因子：
3.7
通讯作者：
Wang Gang
中科院分区：
文献类型：
--
作者：
Gao Jianqing;Hu Yulan;Li Ni;Zhu Xiumei;Li Haijie;Han Min;Miao Peihong;Hu Zhongjie;Wang Gang
目的利用非病毒载体提高基因工程大鼠间充质干细胞（MSCs）的表达水平并延长其表达时间。方法建立一种基于反向转染方法和三维支架的新型转染系统。与常规方法相比，对反向基因转染系统的转染效率进行了评估。引入胶原蛋白海绵和聚对苯二甲酸乙二醇酯无纺布作为支架，反向转染进行三维培养。将编码tgf - β-1的pDNA传递到MSCs中，利用3D非病毒反向转染系统评估其诱导软骨形成的能力。结果在血清存在的情况下，反向转染法诱导的转基因水平高于常规转染法。阴离子明胶的电荷通过影响基因复合物的释放模式和通过血清蛋白在底物上的吸附在该系统中起重要作用。在长时间的体外培养过程中，在3D支架上培养的MSCs比在二维基质上培养和转染的MSCs表现出更高的转基因表达水平和更持久的转基因表达。结论逆转染系统与3D细胞培养支架相结合有利于MSCs的细胞增殖和长时间基因转染。
PurposeTo enhance the level and prolong the duration of gene expression for gene-engineered rat mesenchymal stem cells (MSCs) using non-viral vector.MethodsA novel transfection system based on reverse transfection method and three-dimensional (3D) scaffold was developed. The reverse gene transfection system was evaluated for transfection efficiency compared to conventional methods. Collagen sponge and polyethylene terephthalate non-woven fabric were introduced as scaffolds to perform 3D culture with reverse transfection. pDNA coding TGFβ-1 was delivered to MSCs to assess its ability in inducing chondrogenesis with the 3D non-viral reverse transfection system.ResultsThe reverse transfection method induced higher transgene levels than the conventional transfection in the presence of serum. The electric charge of the anionic gelatin plays an important role in this system by affecting the release pattern of the gene complexes and through the adsorption of serum protein to the substrate. During a long-timein vitroculture, MSCs cultured on 3D scaffolds exhibited a higher transgene expression level and more sustained transgene expression than those cultured and transfected on the two-dimensional substrate.ConclusionsThe combination of reverse transfection system with 3D cell culture scaffold benefits the cell proliferation and long-time gene transfection of MSCs.
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影响因子：
3.8
作者：
Fernandes TG;Kwon SJ;Bale SS;Lee MY;Diogo MM;Clark DS;Cabral JM;Dordick JS
通讯作者：
Dordick JS
影响因子：
14
作者：
Naoki Sakaguchi;C. Kojima;Atsushi Harada;Kazunori Koiwai;K. Shimizu;N. Emi;K. Kono
通讯作者：
Naoki Sakaguchi;C. Kojima;Atsushi Harada;Kazunori Koiwai;K. Shimizu;N. Emi;K. Kono
DOI：
10.1073/pnas.0903269107
发表时间：
2010-03-16
影响因子：
11.1
作者：
Kilian, Kristopher A.;Bugarija, Branimir;Mrksich, Milan
通讯作者：
Mrksich, Milan
影响因子：
56.9
作者：
Cukierman, E;Pankov, R;Yamada, KM
通讯作者：
Yamada, KM
影响因子：
3.8
作者：
Bengali, Z;Pannier, AK;Shea, LD
通讯作者：
Shea, LD