Plasmid DNA transfection using magnetite cationic liposomes for construction of multilayered gene-engineered cell sheet
✍ Scribed by Kosuke Ino; Tamayo Kawasumi; Akira Ito; Hiroyuki Honda
- Publisher
- John Wiley and Sons
- Year
- 2008
- Tongue
- English
- Weight
- 481 KB
- Volume
- 100
- Category
- Article
- ISSN
- 0006-3592
No coin nor oath required. For personal study only.
✦ Synopsis
Abstract
Modification of cellular functions by overexpression of genes is being increasingly practiced for tissue engineering. In the present study, we investigated whether transfection efficiency could be enhanced by magnetofection that involves the use of plasmid DNA (pDNA)/magnetite cationic liposomes (MCLs) complexes (pDNA/MCL) and magnetic force. The transfection efficiencies of the magnetofection technique by pDNA/MCL in fibroblasts and keratinocytes using reporter genes were 36‐ and 10‐fold higher, respectively, than those of a lipofection technique by cationic liposomes. Moreover, in vitro construction of three‐dimensional (3D) tissues is an important challenge. We recently proposed a novel technique termed “magnetic force‐based tissue engineering” (Mag‐TE) to produce 3D tissues. Since the fibroblasts after magnetofection incorporated both magnetite nanoparticles and pDNA, we investigated whether multilayered heterotypic cell sheets expressing transgene could be fabricated by Mag‐TE. First, the fibroblasts were seeded onto an ultra‐low attachment culture plate. When a magnet was placed under the plate, the cells accumulated at the bottom of the culture plate. After 24 h of culture, the transgene‐expressing cells formed a multilayered cell sheet‐like structure. These results indicated that MCLs are a potent biomanipulation tool for both gene transfer and 3D tissue construction, suggesting that these techniques are useful for tissue engineering. Biotechnol. Bioeng. 2008;100: 168–176. © 2007 Wiley Periodicals, Inc.