caa a22 4500 445312408 CHVBK 20180317142634.0 cr unu---uuuuu 170323e20110701xx s 000 0 eng 10.1007/s11095-011-0390-0 doi (NATIONALLICENCE)springer-10.1007/s11095-011-0390-0 Effective Gene Delivery to Mesenchymal Stem Cells Based on the Reverse Transfection and Three-Dimensional Cell Culture System [Elektronische Daten] [Cai-Xia He, Ni Li, Yu-Lan Hu, Xiu-Mei Zhu, Hai-Jie Li, Min Han, Pei-Hong Miao, Zhong-Jie Hu, Gang Wang, Wen-Quan Liang, Yasuhiko Tabata, Jian-Qing Gao] 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. Springer Science+Business Media, LLC, 2011 gene transfection nationallicence mesenchymal stem cells nationallicence non-viral gene vector nationallicence reverse transfection nationallicence three dimensional nationallicence BCA : bicinchonic acid nationallicence CDI : N, N'—carbonyldiimidazole nationallicence CPZ : chlorpromazine nationallicence DMEM : Dulbecco's modified Eagle's medium nationallicence MSCs : mesenchymal stem cells nationallicence PEI : poly (ethylene-imine) nationallicence PET : polyethylene terephthalate nationallicence TGFβ-1 : transforming growth factor β-1 nationallicence He Cai-Xia Institute of Pharmaceutics, College of Pharmaceutical Sciences, Zhejiang University, 388 Yuhangtang Road, 310058, Hangzhou, People's Republic of China aut Li Ni Institute of Pharmaceutics, College of Pharmaceutical Sciences, Zhejiang University, 388 Yuhangtang Road, 310058, Hangzhou, People's Republic of China aut Hu Yu-Lan Institute of Pharmaceutics, College of Pharmaceutical Sciences, Zhejiang University, 388 Yuhangtang Road, 310058, Hangzhou, People's Republic of China aut Zhu Xiu-Mei Institute of Pharmaceutics, College of Pharmaceutical Sciences, Zhejiang University, 388 Yuhangtang Road, 310058, Hangzhou, People's Republic of China aut Li Hai-Jie Institute of Pharmaceutics, College of Pharmaceutical Sciences, Zhejiang University, 388 Yuhangtang Road, 310058, Hangzhou, People's Republic of China aut Han Min Institute of Pharmaceutics, College of Pharmaceutical Sciences, Zhejiang University, 388 Yuhangtang Road, 310058, Hangzhou, People's Republic of China aut Miao Pei-Hong Department of Pharmacy, Zhejiang Provincial Corps Hospital of Chinese People's Armed Police Forces, 314000, Jiaxing, People's Republic of China aut Hu Zhong-Jie Department of Pharmacy, Zhejiang Provincial Corps Hospital of Chinese People's Armed Police Forces, 314000, Jiaxing, People's Republic of China aut Wang Gang Department of Pharmacy, First People's Hospital of Hangzhou, 310006, Hangzhou, People's Republic of China aut Liang Wen-Quan Institute of Pharmaceutics, College of Pharmaceutical Sciences, Zhejiang University, 388 Yuhangtang Road, 310058, Hangzhou, People's Republic of China aut Tabata Yasuhiko Department of Biomaterials, Field of Tissue Engineering Institute for Frontier Medical Sciences, Kyoto University, 53 Kawanara-cho, Shogoin, Sakyo-ku, 606-8507, Kyoto, Japan aut Gao Jian-Qing Institute of Pharmaceutics, College of Pharmaceutical Sciences, Zhejiang University, 388 Yuhangtang Road, 310058, Hangzhou, People's Republic of China aut Pharmaceutical Research Springer US; http://www.springer-ny.com 28/7(2011-07-01), 1577-1590 0724-8741 28:7<1577 2011 28 11095 https://doi.org/10.1007/s11095-011-0390-0 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s11095-011-0390-0 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- He Cai-Xia Institute of Pharmaceutics, College of Pharmaceutical Sciences, Zhejiang University, 388 Yuhangtang Road, 310058, Hangzhou, People's Republic of China aut NATIONALLICENCE 700 1- Li Ni Institute of Pharmaceutics, College of Pharmaceutical Sciences, Zhejiang University, 388 Yuhangtang Road, 310058, Hangzhou, People's Republic of China aut NATIONALLICENCE 700 1- Hu Yu-Lan Institute of Pharmaceutics, College of Pharmaceutical Sciences, Zhejiang University, 388 Yuhangtang Road, 310058, Hangzhou, People's Republic of China aut NATIONALLICENCE 700 1- Zhu Xiu-Mei Institute of Pharmaceutics, College of Pharmaceutical Sciences, Zhejiang University, 388 Yuhangtang Road, 310058, Hangzhou, People's Republic of China aut NATIONALLICENCE 700 1- Li Hai-Jie Institute of Pharmaceutics, College of Pharmaceutical Sciences, Zhejiang University, 388 Yuhangtang Road, 310058, Hangzhou, People's Republic of China aut NATIONALLICENCE 700 1- Han Min Institute of Pharmaceutics, College of Pharmaceutical Sciences, Zhejiang University, 388 Yuhangtang Road, 310058, Hangzhou, People's Republic of China aut NATIONALLICENCE 700 1- Miao Pei-Hong Department of Pharmacy, Zhejiang Provincial Corps Hospital of Chinese People's Armed Police Forces, 314000, Jiaxing, People's Republic of China aut NATIONALLICENCE 700 1- Hu Zhong-Jie Department of Pharmacy, Zhejiang Provincial Corps Hospital of Chinese People's Armed Police Forces, 314000, Jiaxing, People's Republic of China aut NATIONALLICENCE 700 1- Wang Gang Department of Pharmacy, First People's Hospital of Hangzhou, 310006, Hangzhou, People's Republic of China aut NATIONALLICENCE 700 1- Liang Wen-Quan Institute of Pharmaceutics, College of Pharmaceutical Sciences, Zhejiang University, 388 Yuhangtang Road, 310058, Hangzhou, People's Republic of China aut NATIONALLICENCE 700 1- Tabata Yasuhiko Department of Biomaterials, Field of Tissue Engineering Institute for Frontier Medical Sciences, Kyoto University, 53 Kawanara-cho, Shogoin, Sakyo-ku, 606-8507, Kyoto, Japan aut NATIONALLICENCE 700 1- Gao Jian-Qing Institute of Pharmaceutics, College of Pharmaceutical Sciences, Zhejiang University, 388 Yuhangtang Road, 310058, Hangzhou, People's Republic of China aut NATIONALLICENCE 773 0- Pharmaceutical Research Springer US; http://www.springer-ny.com 28/7(2011-07-01), 1577-1590 0724-8741 28:7<1577 2011 28 11095 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer