Dynamic and Label-Free Cell-Based Assays Using the Real-Time Cell Electronic Sensing System

Abstract

Cell-based assays have become an integral part of the preclinical drug development process. Recently, noninvasive label-free cell-based assay technologies have taken center stage, offering important and distinct advantages over and in addition to traditional label-based endpoint assays. Dynamic monitoring of live cells, the preclusion of label, and kinetics are some of the fundamental features of cell-based label-free technologies. In this article we will discuss the real-time cell electronic sensing (RT-CES, ACEA Biosciences Inc., San Diego, CA) system and some of its key applications for cell-based assays such as cell proliferation and cytotoxicity, functional assays for receptor-ligand analysis, cell adhesion and spreading assays, dynamic monitoring of endothelial barrier function, and dynamic monitoring of cell migration and invasion. Also, where appropriate we will briefly discuss other label-free technologies in an application-specific manner.

Isoform-specific differences between Rap1A and Rap1B GTPases in the formation of endothelial cell junctions

“…50,51 The RTCA (Real-Time Cell Analysis) system is an automated real-time method to monitor the impedance generated by a monolayer of cells grown on microelectrode sensors. ECs were preinfected with indicated shRNAs, trypsinized and replated at confluent cell density onto microelectrode coated dishes and analyzed for 24 hrs.…”

“…Changes in impedance reflect changes in barrier function and permeability. 50 HUVEC preinfected with the indicated shRNA for 48-72 hrs were counted pDONR221 (also from Invitrogen, 12536017) by Gateway BP recombination and then into pAd-CMV-Dest Gateway vector by LR recombination.…”

Quantification of Cell Migration and Invasion, and Their Association with Periostin in Anaplastic Thyroid Cancer, Using a Real-time Cell Analyzer

“…This system uses impedance detection to produce a cell index for the continuous monitoring of cell viability, migration, and invasion 6 . Previously, we reported on the migration and invasion of oral squamous cell carcinomas OSCCs using the RTCA system 5 .…”

Digital Holography and Cell Studies

“…It is a real-time cell analyzer that measures living cells without labeling. Electric impedance allows measurements of cellular processes such as proliferation, cytotoxicity, invasion, migration and cell viability (Atienza et al, 2006;Boyd et al, 2008;Ge et al, 2009) and give results comparable to imaging methods in some ways. Different staining methods have been developed that enable cells to absorb light.…”