Raz-Ben Aroush Dikla, Yehudai-Resheff Shlomit, Keren Kinneret
Department of Physics, Technion- Israel Institute of Technology, Haifa, Israel.
Department of Physics, Technion- Israel Institute of Technology, Haifa, Israel; Department of Hematology, Rambam Healthcare Campus, Haifa, Israel.
Methods Cell Biol. 2015;125:409-22. doi: 10.1016/bs.mcb.2014.11.005. Epub 2015 Jan 8.
We describe a method for electroporation-induced fusion of giant unilamellar vesicles (GUVs) with the plasma membrane of adherent cells. Using this method, the area of the cell membrane can be abruptly increased and various lipids can be introduced into the membrane. The process involves two steps: (1) the formation of GUVs with controlled membrane composition and (2) electrofusion of the GUVs to living cells using an electroporator for adherent cells. We demonstrate the technique on fish epithelial keratocytes and human foreskin fibroblasts, and discuss the influence of the composition of the GUVs on the fusion process.
我们描述了一种通过电穿孔诱导巨型单层囊泡(GUVs)与贴壁细胞的质膜融合的方法。使用这种方法,可以突然增加细胞膜的面积,并将各种脂质引入膜中。该过程包括两个步骤:(1)形成具有可控膜组成的GUVs,以及(2)使用用于贴壁细胞的电穿孔仪将GUVs与活细胞进行电融合。我们在鱼类上皮角膜细胞和人包皮成纤维细胞上展示了该技术,并讨论了GUVs的组成对融合过程的影响。