title: Cell electrofusion using nanosecond electric pulses.
abstract: Electrofusion is an efficient method for fusing cells using short-duration high-voltage electric pulses. However, electrofusion yields are very low when fusion partner cells differ considerably in their size, since the extent of electroporation (consequently membrane fusogenic state) with conventionally used microsecond pulses depends proportionally on the cell radius. We here propose a new and innovative approach to fuse cells with shorter, nanosecond (ns) pulses. Using numerical calculations we demonstrate that ns pulses can induce selective electroporation of the contact areas between cells (i.e. the target areas), regardless of the cell size. We then confirm experimentally on B16-F1 and CHO cell lines that electrofusion of cells with either equal or different size by using ns pulses is indeed feasible. Based on our results we expect that ns pulses can improve fusion yields in electrofusion of cells with different size, such as myeloma cells and B lymphocytes in hybridoma technology.
authors.affiliation: 1] University of Ljubljana, Faculty of Electrical Engineering, Tržaška cesta 25, SI-1000 Ljubljana, Slovenia [2].
authors.forename: Lea
authors.initials: L
authors.lastname: Rems
authors.forename: Marko
authors.initials: M
authors.lastname: Ušaj
authors.forename: Maša
authors.initials: M
authors.lastname: Kandušer
authors.forename: Matej
authors.initials: M
authors.lastname: Reberšek
authors.forename: Damijan
authors.initials: D
authors.lastname: Miklavčič
authors.forename: Gorazd
authors.initials: G
authors.lastname: Pucihar
mesh_terms: D000818:Animals; D016466:CHO Cells; D002459:Cell Fusion / Q000379:methods*; D048429:Cell Size; D003412:Cricetulus; D004560:Electricity; D018274:Electroporation / Q000379:methods*; D006825:Hybridomas / Q000166:cytology; D008546:Melanoma, Experimental; D051379:Mice; D011674:Pulse / Q000379:methods*