title: Optical reprogramming of human cells in an ultrashort femtosecond laser microfluidic transfection platform.
abstract: Induced pluripotent stem cell (iPS cell) technology can be used to produce unlimited numbers of functional cells for both research and therapeutic purposes without ethical controversy. Typically, viruses are applied for efficient intracellular delivery of genes/transcription factors to generate iPS cells. However, the viral genomic integration may cause a risk of mutation as well as tumor formation therefore limits its clinical application. Here we demonstrate that spatially shaped extreme ultrashort laser pulses of sub-20 femtoseconds induce transient membrane permeabilisation which enables contamination-free transfection of cells in a microfluidic tube with multiple genes at the individual cell level in order to achieve optical reprogramming of large cell populations. We found that the ultrashort femtosecond laser-microfluidic cell transfection platform enhanced the efficacy of iPS-like colony-forming following merely a single transfection. Illustration of the spatially shaped femtosecond laser-assisted microfluidic cell transfection platform for production of iPS cell colonies.
authors.affiliation: Saarland University, Department of Biophotonics and Laser Technology, Campus A5.1, 66123, Saarbrücken, Germany. a.uchugonova@blt.uni-saarland.de.|JenLab GmbH, Schillerstr. 1, 07745, Jena, Germany. a.uchugonova@blt.uni-saarland.de.
authors.forename: Aisada
authors.initials: A
authors.lastname: Uchugonova
authors.affiliation: Saarland University, Department of Biophotonics and Laser Technology, Campus A5.1, 66123, Saarbrücken, Germany.|JenLab GmbH, Schillerstr. 1, 07745, Jena, Germany.
authors.forename: Hans Georg
authors.initials: HG
authors.lastname: Breunig
authors.affiliation: Saarland University, Department of Biophotonics and Laser Technology, Campus A5.1, 66123, Saarbrücken, Germany.
authors.forename: Ana
authors.initials: A
authors.lastname: Batista
authors.affiliation: Saarland University, Department of Biophotonics and Laser Technology, Campus A5.1, 66123, Saarbrücken, Germany.|JenLab GmbH, Schillerstr. 1, 07745, Jena, Germany.
authors.forename: Karsten
authors.initials: K
authors.lastname: König
mesh_terms: D002478:Cells, Cultured; D065150:Cellular Reprogramming; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D007834:Lasers; D044085:Microfluidics / Q000379:methods*; D014162:Transfection / Q000379:methods*
keywords: femtosecond laser; flow cell; induced pluripotent stem cells; optical transfection; reprogramming