Yang T A, Heiser W C, Sedivy J M
Department of Molecular Biophysics and Biochemistry, Yale University School of Medicine, New Haven, CT 06520, USA.
Nucleic Acids Res. 1995 Aug 11;23(15):2803-10. doi: 10.1093/nar/23.15.2803.
Electroporation is a common technique for the introduction of DNA molecules into living cells. The method is currently limited by the necessity of applying the electrical discharge to cells in suspension. Adherent cells must therefore be removed from their substratum, which can induce unwanted physiological effects. We report here a new procedure for in situ electroporation of cells grown on microporous membranes of polyethylene terephthalate (PET) or polyester (PE). We demonstrate that this method of in situ electroporation employs only readily available materials and standard electroporation devices without any modifications, is as efficient as conventional electroporation of cells in suspension, and is applicable to a wide range of cell types. Efficient electroporation can be achieved under conditions of minimal cell killing, and can be performed with quiescent cells as well as with confluent epithelial sheets. The method is a useful extension of electroporation technology, and will allow the application of electroporation to a wider spectrum of biological systems.
电穿孔是一种将DNA分子导入活细胞的常用技术。该方法目前受限于必须对悬浮细胞施加放电。因此，贴壁细胞必须从其基质上移除，这可能会诱导不必要的生理效应。我们在此报告一种对生长在聚对苯二甲酸乙二酯（PET）或聚酯（PE）微孔膜上的细胞进行原位电穿孔的新方法。我们证明，这种原位电穿孔方法仅使用现成的材料和标准电穿孔设备，无需任何修改，与悬浮细胞的传统电穿孔一样高效，并且适用于广泛的细胞类型。在最小细胞杀伤的条件下可以实现高效电穿孔，并且可以对静止细胞以及汇合的上皮细胞片进行电穿孔。该方法是电穿孔技术的有益扩展，将使电穿孔能够应用于更广泛的生物系统。
