title: Biophysical effects in off-resonant gold nanoparticle mediated (GNOME) laser transfection of cell lines, primary- and stem cells using fs laser pulses.
abstract: Gold nanoparticle mediated (GNOME) laser transfection is a powerful technique to deliver small biologically relevant molecules into cells. However, the transfection of larger and especially negatively charged DNA remains challenging. The efficiency for pDNA was 0.57% using parameter that does not influence the endo- and exogenous DNA. In order to gain a deeper understanding of the actual molecule uptake process, the uptake efficiency was determined using molecules of different sizes. It was evaluated that uncharged dextran molecules (2000 kDa) were delivered with an efficiency of 68%. The intracellular distribution of injected molecules was visualized and larger molecules were primary found in the cytoplasm. Patch clamp measurements suggested a permeabilization time up to 15 minutes. The uptake efficiency depended on the size and charge of the molecule to deliver as well as the cell size. A minor role for transfection plays the cell type since primary stem cells were successfully transfected. The perforation efficiency of semi-adherent and suspension cells is influenced by the cell and molecule size.
authors.affiliation: Biomedical Optics Department, Laser Zentrum Hannover e. V., Hollerithallee 8, 30419, Hannover, Germany. m.schomaker@lzh.de.
authors.forename: Markus
authors.initials: M
authors.lastname: Schomaker
authors.affiliation: Department Hematology, Oncology and Palliative Medicine, University of Rostock, Ernst-Heydemann-Str. 6, 18057, Rostock, Germany.
authors.forename: Doreen
authors.initials: D
authors.lastname: Killian
authors.affiliation: Small Animal Clinic, University of Veterinary Medicine Hannover, Bünteweg 9, 30559, Hannover, Germany.
authors.forename: Saskia
authors.initials: S
authors.lastname: Willenbrock
authors.affiliation: Biomedical Optics Department, Laser Zentrum Hannover e. V., Hollerithallee 8, 30419, Hannover, Germany.
authors.forename: Dag
authors.initials: D
authors.lastname: Heinemann
authors.affiliation: Biomedical Optics Department, Laser Zentrum Hannover e. V., Hollerithallee 8, 30419, Hannover, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Kalies
authors.affiliation: Institute of Biophysics, Leibniz University Hannover, Herrenhaeuserstr. 2, 30419, Hannover, Germany.
authors.forename: Anaclet
authors.initials: A
authors.lastname: Ngezahayo
authors.affiliation: Small Animal Clinic, University of Veterinary Medicine Hannover, Bünteweg 9, 30559, Hannover, Germany.
authors.forename: Ingo
authors.initials: I
authors.lastname: Nolte
authors.affiliation: Biomedical Optics Department, Laser Zentrum Hannover e. V., Hollerithallee 8, 30419, Hannover, Germany.
authors.forename: Tammo
authors.initials: T
authors.lastname: Ripken
authors.affiliation: Department Hematology, Oncology and Palliative Medicine, University of Rostock, Ernst-Heydemann-Str. 6, 18057, Rostock, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Junghanß
authors.affiliation: Biomedical Optics Department, Laser Zentrum Hannover e. V., Hollerithallee 8, 30419, Hannover, Germany.|Department of Cardiothoracic Transplantation and Vascular Surgery, Hannover Medical School, Carl-Neuberg-Str. 1, 30625, Hannover, Germany.
authors.forename: Heiko
authors.initials: H
authors.lastname: Meyer
authors.affiliation: Department Hematology, Oncology and Palliative Medicine, University of Rostock, Ernst-Heydemann-Str. 6, 18057, Rostock, Germany.|Small Animal Clinic, University of Veterinary Medicine Hannover, Bünteweg 9, 30559, Hannover, Germany.
authors.forename: Hugo
authors.initials: H
authors.lastname: Murua Escobar
authors.affiliation: Biomedical Optics Department, Laser Zentrum Hannover e. V., Hollerithallee 8, 30419, Hannover, Germany.|Institut für Quantenoptik, Leibniz Universität Hannover, Welfengarten 1, 30167, Hannover, Germany.
authors.forename: Alexander
authors.initials: A
authors.lastname: Heisterkamp
mesh_terms: D000818:Animals; D055592:Biophysical Phenomena; D045744:Cell Line, Tumor; D002463:Cell Membrane Permeability; D048429:Cell Size; D004247:DNA / Q000235:genetics / Q000378:metabolism; D004285:Dogs; D006046:Gold / Q000737:chemistry*; D007834:Lasers; D053768:Metal Nanoparticles / Q000737:chemistry*; D013234:Stem Cells / Q000378:metabolism*; D014162:Transfection / Q000379:methods*
chemical_list: D006046:Gold; D004247:DNA
keywords: DNA; GNOME laser transfection; gold nanoparticles; molecule size; stem cells