title: Delivery of episomal vectors into primary cells by means of commercial transfection reagents.
abstract: Although episomal vectors are commonly transported into cells by electroporation, a number of electroporation-derived problems have led to the search for alternative transfection protocols, such as the use of transfection reagents, which are inexpensive and easy to handle. Polyplex-mediated transport of episomal vectors into the cytoplasm has been conducted successfully in immortalized cell lines, but no report exists of successful transfection of primary cells using this method. Accordingly, we sought to optimize the conditions for polyplex-mediated transfection for effective delivery of episomal vectors into the cytoplasm of primary mouse embryonic fibroblasts. Episomal vectors were complexed with the commercially available transfection reagents Lipofectamine 2000, FuGEND HD and jetPEI. The ratio of transfection reagent to episomal vectors was varied, and the subsequent transfection efficiency and cytotoxicity of the complexes were analyzed using flow cytometry and trypan blue exclusion assay, respectively. No cytotoxicity and the highest transfection yield were observed when the ratio of transfection reagent to episomal vector was 4 (v/wt) in the cases of Lipofectamine 2000 and FuGENE HD, and 2 in the case of jetPEI. Of the three transfection reagents tested, jetPEI showed the highest transfection efficiency without any cytotoxicity. Thus, we confirmed that the transfection reagent jetPEI could be used to effectively deliver episomal vectors into primary cells without electroporation.
authors.affiliation: Department of Animal Life Science, Kangwon National University, Chuncheon 200-701, South Korea.
authors.forename: Na Rae
authors.initials: NR
authors.lastname: Han
authors.affiliation: Department of Animal Life Science, Kangwon National University, Chuncheon 200-701, South Korea.
authors.forename: Hyun
authors.initials: H
authors.lastname: Lee
authors.affiliation: Department of Animal Life Science, Kangwon National University, Chuncheon 200-701, South Korea.
authors.forename: Song
authors.initials: S
authors.lastname: Baek
authors.affiliation: Division of Animal Resource Science, Kangwon National University, Chuncheon 200-701, South Korea.
authors.forename: Jung Im
authors.initials: JI
authors.lastname: Yun
authors.affiliation: Department of Animal Life Science, Kangwon National University, Chuncheon 200-701, South Korea; Division of Animal Resource Science, Kangwon National University, Chuncheon 200-701, South Korea.
authors.forename: Kyu Hyun
authors.initials: KH
authors.lastname: Park
authors.affiliation: Department of Animal Life Science, Kangwon National University, Chuncheon 200-701, South Korea; Division of Applied Animal Science, Kangwon National University, Chuncheon 200-701, South Korea. Electronic address: stlee76@kangwon.ac.kr.
authors.forename: Seung Tae
authors.initials: ST
authors.lastname: Lee
mesh_terms: D000818:Animals; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D018274:Electroporation / Q000379:methods; D005822:Genetic Vectors / Q000008:administration & dosage* / Q000737:chemistry / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000235:genetics; D008055:Lipids / Q000737:chemistry / Q000633:toxicity; D051379:Mice; D010957:Plasmids / Q000008:administration & dosage* / Q000737:chemistry / Q000235:genetics; D014162:Transfection / Q000379:methods*
chemical_list: C411955:FuGene; D008055:Lipids; C086724:Lipofectamine; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins
keywords: Delivery; Electroporation; Episomal vector; Primary cells; Transfection reagent