基于 ECIS 的电围栏方法用于测量不同基底上的人角质形成细胞的迁移。

ECIS Based Electric Fence Method for Measurement of Human Keratinocyte Migration on Different Substrates.

机构信息

Department of Biomedical Engineering, National Yang Ming Chiao Tung University, Taipei 112, Taiwan.

Department of Aesthetic Medicine, Chen Hsin General Hospital, Taipei 112, Taiwan.

出版信息

Biosensors (Basel). 2022 May 3;12(5):293. doi: 10.3390/bios12050293.

DOI:10.3390/bios12050293

PMID:35624596

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC9138674/

Abstract

Electric Cell-substrate Impedance Sensing (ECIS) is an impedance-based, real-time, and label-free measuring system for monitoring cellular activities in tissue culture. Previously, ECIS wound healing assay has been used to wound cells with high electric current and monitor the subsequent cell migration. In this study, we applied ECIS electric fence (EF) method, an alternative to electrical wounding, to assess the effects of different surface coatings on human keratinocyte (HaCaT) migration. The EF prevents inoculated cells from attaching or migrating to the fenced electrode surface while maintaining the integrity of the surface coating. After the EF is turned off, cells migrate into the cell-free area, and the increase in measured impedance is monitored. We cultured HaCaT cells on gold electrodes without coating or coated with poly-L-lysin (PLL), poly-D-lysine (PDL), or type-I collagen. We quantified migration rates according to the different slopes in the impedance time series. It was observed that either poly-L-lysine (PLL) or poly-D-lysine (PDL) limits cell adhesion and migration rates. Furthermore, the surface charge of the coated substrate in the culture condition positively correlates with the cell adhesion and migration process. Our results indicate that the EF method is useful for determining cell migration rates on specific surface coatings.

摘要

电细胞-基质阻抗传感（ECIS）是一种基于阻抗的实时、无标记测量系统，用于监测组织培养中的细胞活性。此前，ECIS 伤口愈合分析已被用于用高电流损伤细胞，并监测随后的细胞迁移。在这项研究中，我们应用了 ECIS 电围栏（EF）方法，这是一种替代电损伤的方法，来评估不同表面涂层对人角质形成细胞（HaCaT）迁移的影响。EF 可防止接种的细胞附着或迁移到围栏电极表面，同时保持表面涂层的完整性。EF 关闭后，细胞迁移到无细胞区域，并且监测测量阻抗的增加。我们在没有涂层或涂有聚-L-赖氨酸（PLL）、聚-D-赖氨酸（PDL）或 I 型胶原的金电极上培养 HaCaT 细胞。我们根据阻抗时间序列中的不同斜率来量化迁移率。结果表明，聚-L-赖氨酸（PLL）或聚-D-赖氨酸（PDL）均限制细胞的粘附和迁移率。此外，在培养条件下涂层基底的表面电荷与细胞粘附和迁移过程呈正相关。我们的结果表明，EF 方法可用于确定特定表面涂层上的细胞迁移率。