title: Optimization of 25 kDa linear polyethylenimine for efficient gene delivery.
abstract: A 25-kDa linear polyethylenimine (25 kDa L-PEI) has proven to be efficient and versatile agent for gene delivery. Therefore, we determined the optimal transfection conditions of 25 kDa L-PEI and examined whether it has comparable transfection efficiency with other commercially available reagents, ExGen 500, LipofectAMINE 2000, and Effectene by using EGFP expression vector in different cell lines. Transfection efficiency and cytotoxicity were measured by flow cytometry. First of all, we determined the optimal ratio of nitrogen to phosphorous (N/P) and DNA concentration. With the increase of N/P ratio and DNA amounts, transfection efficiency increased with a slight variation in cell types. The optimal amounts of 25 kDa L-PEI were determined at N/P ratio 40 and DNA concentration varied among the cell types. In addition, 25 kDa L-PEI worked efficiently and was less toxic than other reagents. However, the efficiency and toxicity of all these reagents varied according to cell types as well as the ratio of DNA to reagents and the amounts of DNA. Our finding illustrates the importance of optimal transfection conditions of 25 kDa L-PEI to obtain maximal transgene expression with less cytotoxicity. Importantly, the optimization of those conditions may make possible to perform transfection cost-effectively and efficiently.
authors.affiliation: CHA Stem Cell Institute, Graduate School of Life Science and Biotechnology, Pochon CHA University, Kangnam-Gu, Seoul 135-907, South Korea.
authors.forename: Sun-Hyung
authors.initials: SH
authors.lastname: Huh
authors.forename: Hyun-Jin
authors.initials: HJ
authors.lastname: Do
authors.forename: Hye-Young
authors.initials: HY
authors.lastname: Lim
authors.forename: Dong-Ku
authors.initials: DK
authors.lastname: Kim
authors.forename: Seong-Jun
authors.initials: SJ
authors.lastname: Choi
authors.forename: Hyuk
authors.initials: H
authors.lastname: Song
authors.forename: Nam-Hyung
authors.initials: NH
authors.lastname: Kim
authors.forename: Jin-Ki
authors.initials: JK
authors.lastname: Park
authors.forename: Won-Kyong
authors.initials: WK
authors.lastname: Chang
authors.forename: Hyung-Min
authors.initials: HM
authors.lastname: Chung
authors.forename: Jae-Hwan
authors.initials: JH
authors.lastname: Kim
mesh_terms: D000818:Animals; D019556:COS Cells; D002460:Cell Line; D002522:Chlorocebus aethiops; D004274:DNA, Recombinant / Q000008:administration & dosage / Q000235:genetics; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D008970:Molecular Weight; D009584:Nitrogen / Q000032:analysis; D010758:Phosphorus / Q000032:analysis; D011094:Polyethyleneimine / Q000737:chemistry* / Q000633:toxicity; D011994:Recombinant Proteins / Q000235:genetics; D014162:Transfection
chemical_list: D004274:DNA, Recombinant; D011994:Recombinant Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D010758:Phosphorus; D011094:Polyethyleneimine; D009584:Nitrogen