Brooks Justin R, Heiman Tyler C, Lorenzen Sawyer R, Mungloo Ikhlaas, Mirfendereski Siamak, Park Jae Sung, Yang Ruiguo
Department of Mechanical and Materials Engineering, University of Nebraska-Lincoln, Lincoln, NE, 68588, USA.
Nebraska Center for Integrated Biomolecular Communication, University of Nebraska-Lincoln, Lincoln, NE, 68588, USA.
Small. 2024 Jun;20(25):e2310221. doi: 10.1002/smll.202310221. Epub 2024 Feb 23.
Porous substrate electroporation (PSEP) is a promising new method for intracellular delivery, yet fundamentals of PSEP are not well understood, especially the intermediate processes leading to delivery. PSEP is an electrical method, yet the relationship between PSEP and electrical impedance remains underexplored. In this study, a device capable of measuring impedance and performing PSEP is developed and the changes in transepithelial electrical impedance (TEEI) are monitored. These measurements show TEEI increases following PSEP, unlike other electroporation methods. The authors then demonstrate how cell culture conditions and electrical waveforms influence this response. More importantly, TEEI response features are correlated with viability and delivery efficiency, allowing prediction of outcomes without fluorescent cargo, imaging, or image processing. This label-free delivery also allows improved temporal resolution of transient processes following PSEP, which the authors expect will aid PSEP optimization for new cell types and cargos.
多孔基质电穿孔(PSEP)是一种很有前景的细胞内递送新方法,但PSEP的基本原理尚未得到充分理解,尤其是导致递送的中间过程。PSEP是一种电方法,但PSEP与电阻抗之间的关系仍未得到充分研究。在本研究中,开发了一种能够测量阻抗并进行PSEP的装置,并监测跨上皮电阻抗(TEEI)的变化。这些测量结果表明,与其他电穿孔方法不同,PSEP后TEEI会增加。作者随后展示了细胞培养条件和电波形如何影响这种反应。更重要的是,TEEI反应特征与活力和递送效率相关,无需荧光货物、成像或图像处理就能预测结果。这种无标记递送还能提高PSEP后瞬态过程的时间分辨率,作者预计这将有助于针对新细胞类型和货物优化PSEP。