title: New non-viral method for gene transfer into primary cells.
abstract: The availability of genetically altered cells is an essential prerequisite for many scientific and therapeutic applications including functional genomics, drug development, and gene therapy. Unfortunately, the efficient gene transfer into primary cells is still problematic. In contrast to transfections of most cell lines, which can be successfully performed using a variety of methods, the introduction of foreign DNA into primary cells requires a careful selection of gene transfer techniques. Whereas viral strategies are time consuming and involve safety risks, non-viral methods proved to be inefficient for most primary cell types. The Nucleofector technology is a novel gene transfer technique designed for primary cells and hard-to-transfect cell lines. This non-viral gene transfer method is based on a cell type specific combination of electrical parameters and solutions. In this report, we show efficient transfer of DNA expression vectors and siRNA oligonucleotides into a variety of primary cell types from different species utilizing the Nucleofector technology, including human B-CLL cells, human CD34+ cells, human lymphocytes, rat cardiomyocytes, human, porcine, and bovine chondrocytes, and rat neurons.
authors.affiliation: amaxa GmbH, Cologne, Germany.
authors.forename: Oliver
authors.initials: O
authors.lastname: Gresch
authors.forename: Felix B
authors.initials: FB
authors.lastname: Engel
authors.forename: Dobrila
authors.initials: D
authors.lastname: Nesic
authors.forename: Thi Thai
authors.initials: TT
authors.lastname: Tran
authors.forename: Hazel M
authors.initials: HM
authors.lastname: England
authors.forename: Emma S
authors.initials: ES
authors.lastname: Hickman
authors.forename: Ida
authors.initials: I
authors.lastname: Körner
authors.forename: Li
authors.initials: L
authors.lastname: Gan
authors.forename: Sylvia
authors.initials: S
authors.lastname: Chen
authors.forename: Susana
authors.initials: S
authors.lastname: Castro-Obregon
authors.forename: Rainer
authors.initials: R
authors.lastname: Hammermann
authors.forename: Jürgen
authors.initials: J
authors.lastname: Wolf
authors.forename: Herbert
authors.initials: H
authors.lastname: Müller-Hartmann
authors.forename: Michael
authors.initials: M
authors.lastname: Nix
authors.forename: Gregor
authors.initials: G
authors.lastname: Siebenkotten
authors.forename: Günter
authors.initials: G
authors.lastname: Kraus
authors.forename: Klaus
authors.initials: K
authors.lastname: Lun
mesh_terms: D000818:Animals; D002318:Cardiovascular Diseases / Q000628:therapy; D002417:Cattle; D002478:Cells, Cultured; D019902:Chondrocytes / Q000378:metabolism; D018274:Electroporation / Q000379:methods*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D008214:Lymphocytes / Q000378:metabolism; D032342:Muscle Cells / Q000378:metabolism; D009474:Neurons / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D051381:Rats; D014162:Transfection / Q000379:methods*; D014780:Viruses / Q000235:genetics
chemical_list: D034741:RNA, Small Interfering