Effective Gene Delivery to Mesenchymal Stem Cells Based on the Reverse Transfection and Three-Dimensional Cell Culture System

• Published in: Pharmaceutical research Vol. 28; no. 7; pp. 1577 - 1590
• Main Authors: He, Cai-Xia, Li, Ni, Hu, Yu-Lan, Zhu, Xiu-Mei, Li, Hai-Jie, Han, Min, Miao, Pei-Hong, Hu, Zhong-Jie, Wang, Gang, Liang, Wen-Quan, Tabata, Yasuhiko, Gao, Jian-Qing
• Format: Journal Article
• Language: English
• Published: Boston Springer US 01.07.2011; Springer; Springer Nature B.V
• Subjects: Animals; Biochemistry; Biological and medical sciences; Biomedical and Life Sciences; Biomedical Engineering and Bioengineering; Biomedicine; Cell culture; Cell Culture Techniques; Cell Differentiation; Cells, Cultured; DNA - genetics; Gene expression; Gene Expression Regulation; Gene Transfer Techniques; General pharmacology; HeLa Cells; Humans; Male; Medical Law; Medical sciences; Mesenchymal Stromal Cells - cytology; Microscopy, Electron, Scanning; Pharmaceutical sciences; Pharmaceutical technology. Pharmaceutical industry; Pharmacology. Drug treatments; Pharmacology/Toxicology; Pharmacy; Rats; Rats, Sprague-Dawley; Research Paper; Stem cells; three dimensional; reverse transfection; mesenchymal stem cells; gene transfection; non-viral gene vector; Pharmaceutical technology; Stem cell; In vitro; Genetic transfer; Three dimensional culture; Transfection; Gene; Genetics; Gene therapy; Vector
• ISSN: 0724-8741; 1573-904X
• DOI: 10.1007/s11095-011-0390-0

ABSTRACT Purpose To enhance the level and prolong the duration of gene expression for gene-engineered rat mesenchymal stem cells (MSCs) using non-viral vector. Methods A 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. Results The 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-time in vitro culture, 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. Conclusions The combination of reverse transfection system with 3D cell culture scaffold benefits the cell proliferation and long-time gene transfection of MSCs.