Department of Stem Cell Biology, State Research Institute, Centre for Innovative Medicine, Santariskiu str. 5, LT-08406 Vilnius, Lithuania.
Laboratory of Bioelectrics, State Research Institute, Center for Physical Sciences and Technology, Sauletekio ave. 3, LT-10257, Vilnius, Lithuania.
Exp Cell Res. 2020 Apr 1;389(1):111877. doi: 10.1016/j.yexcr.2020.111877. Epub 2020 Jan 25.
Reversible electroporation is a temporary permeabilization of cell membrane through the formation of transient pores created by short high voltage electric pulses. This method has numerous applications in biology and biotechnology and has become an important technique in molecular medicine. Reversible electroporation is usually used to transfer macromolecules into the cells. However, the delivery of large molecules such as proteins into cells without loss of cell viability remains a challenge. In our study, we investigated whether electroporation can be used for this purpose. The study was performed with the primary mouse splenocytes and Jurkat cell line. The electroporation efficacy was evaluated by flow cytometry. We used the reversible electroporation for intracellular marker detection investigating antibody and fluorescein-conjugated dextran transfer efficiency, cell viability and metabolic activity. We have found that reversible electroporation parameters can be optimized for efficient transfer of large molecules such as antibodies/proteins into live cells without a significant loss of cell viability. We conclude that a well-established and relatively easy method of reversible electroporation can be adjusted to detect intracellular biomarkers in viable cells. This is a new approach on how electroporation could be utilised in medicine and biological research to detect rare subpopulations of cells that produce specific markers and to keep cells viable. This would allow the use of these rare subpopulations of isolated cells for further research and personalized medicine.
可逆电穿孔是通过短时间的高压电脉冲形成瞬时孔来暂时破坏细胞膜的通透性。这种方法在生物学和生物技术中有许多应用，已成为分子医学中的重要技术。可逆电穿孔通常用于将大分子物质转入细胞内。然而，在不损失细胞活力的情况下将大的分子物质如蛋白质递送到细胞内仍然是一个挑战。在我们的研究中，我们研究了电穿孔是否可以用于此目的。该研究使用原代小鼠脾细胞和 Jurkat 细胞系进行。通过流式细胞术评估电穿孔效果。我们使用可逆电穿孔进行细胞内标记物检测，研究了抗体和荧光素标记的葡聚糖的转移效率、细胞活力和代谢活性。我们发现，可逆电穿孔参数可以进行优化，以有效地将大的分子物质（如抗体/蛋白质）转移到活细胞中，而不会显著降低细胞活力。我们得出结论，一种成熟且相对简单的可逆电穿孔方法可以调整为检测活细胞内的生物标志物。这是一种新的方法，电穿孔可以用于医学和生物学研究，以检测产生特定标志物的稀有细胞亚群，并保持细胞活力。这将允许使用这些分离的稀有细胞亚群进行进一步的研究和个性化医疗。
