title: Manipulation of lipoplex concentration at the cell surface boosts transfection efficiency in hard-to-transfect cells.
abstract: To date, efficiency upon non-viral DNA delivery remains low and this implies the existence of unidentified transfection barriers. Here we explore the mechanisms of action of multicomponent (MC) cationic liposome/DNA complexes (lipoplexes) by a combination of reporter technologies, dynamic light scattering (DLS), synchrotron small angle X-ray scattering (SAXS), fluorescence activated cell sorting (FACS) analysis and laser scanning confocal microscopy (LSCM) in live cells. Lipofectamine - the gold standard among transfection reagents - was used as a reference. On the basis of our results, we suggest that an additional transfection barrier impairs transfection efficiency, that is: low lipoplex concentration at the cell surface. Based on the acquired knowledge we propose an optimized transfection protocol that allowed us to efficiently transfect DND41, JURKAT, MOLT3, P12-ICHIKAWA, ALL-SILL, TALL-1 human T-cell acute lymphoblastic leukemia (T-ALL) cell lines known to be difficult-to-transfect by using non-viral vectors and where LFN-based technologies fail to give satisfactory results.
authors: Department of Molecular Medicine, "Sapienza" University of Rome, Rome, Italy.
authors: Sara
authors: S
authors: Palchetti
authors: Department of Molecular Medicine, "Sapienza" University of Rome, Rome, Italy.
authors: Daniela
authors: D
authors: Pozzi
authors: Department of Bioscience and Biotechnology, University of Camerino, Camerino, MC, Italy.
authors: Cristina
authors: C
authors: Marchini
authors: Department of Bioscience and Biotechnology, University of Camerino, Camerino, MC, Italy.
authors: Augusto
authors: A
authors: Amici
authors: Department of Bioscience and Biotechnology, University of Camerino, Camerino, MC, Italy.
authors: Cristina
authors: C
authors: Andreani
authors: Department of Bioscience and Biotechnology, University of Camerino, Camerino, MC, Italy.
authors: Caterina
authors: C
authors: Bartolacci
authors: Department of Molecular Medicine, "Sapienza" University of Rome, Rome, Italy; Department of Bioscience and Biotechnology, University of Camerino, Camerino, MC, Italy.
authors: Luca
authors: L
authors: Digiacomo
authors: Department of Bioscience and Biotechnology, University of Camerino, Camerino, MC, Italy.
authors: Valentina
authors: V
authors: Gambini
authors: Center for Nanotechnology Innovation @NEST, Istituto Italiano di Tecnologia, Pisa, Italy.
authors: Francesco
authors: F
authors: Cardarelli
authors: Center for Nanotechnology Innovation @NEST, Istituto Italiano di Tecnologia, Pisa, Italy.
authors: Carmine
authors: C
authors: Di Rienzo
authors: Istituto Italiano di Tecnologia, CLNS@Sapienza, Rome, Italy.
authors: Giovanna
authors: G
authors: Peruzzi
authors: Institute of inorganic Chemistry, Graz University of Technology, Graz, Austria.
authors: Heinz
authors: H
authors: Amenitsch
authors: Istituto Italiano di Tecnologia, CLNS@Sapienza, Rome, Italy.
authors: Rocco
authors: R
authors: Palermo
authors: Department of Molecular Medicine, "Sapienza" University of Rome, Rome, Italy; Istituto Pasteur-Fondazione Cenci Bolognetti, Sapienza University of Rome, Rome, Italy.
authors: Isabella
authors: I
authors: Screpanti
authors: Department of Molecular Medicine, "Sapienza" University of Rome, Rome, Italy. Electronic address: giulio.caracciolo@uniroma1.it.
authors: Giulio
authors: G
authors: Caracciolo
mesh_terms: D000818:Animals; D002460:Cell Line; D004247:DNA; D006801:Humans; D008055:Lipids; D008081:Liposomes; D053838:Scattering, Small Angle; D014162:Transfection; D014961:X-Ray Diffraction
chemical_list: D008055:Lipids; D008081:Liposomes; D004247:DNA
keywords: Cationic liposomes; Cell transfection; Hard-to-transfect cells; Lipoplexes; Transfection efficiency