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Label-free monitoring of T cell activation by the impedance-based system

Jun 2, 2017

A variety of in vitro cellular systems are currently employed for analyzing T cell activation, yet all suffer from some combination of low throughput, unnatural conditions and long assay times. Label-free technologies are capable of detecting phenotypic responses to treatments under physiological conditions, thereby potentially accelerating drug discovery by facilitating the use of disease-relevant cell models for functional assessment and clinical diagnosis. The xCELLigence system is an impedance based label-free platform that allows for dynamic monitoring of subtle morphological and adhesive changes in cells, such as those induced during T cell activation. Here we describe the development and validation of a T cell activation assay based upon electrical impedance. Co-activation of Jurkat cells with anti-CD28 and anti-CD3 functional antibodies led to impedance changes that were rapidly and sensitively recorded (within 30 minutes). This phenomenon was also observed in human peripheral blood mononuclear cells. These changes reflect morphological and adhesive alterations correlated with cytoskeletal reorganization as verified by microscopy. They were functionally dependent on canonical T cell signaling pathways, including calcium-mediated signals and Src family kinases because relevant inhibitors impaired T cell activation. Our results provide a convenient approach to measure T cell activation in real-time and to elucidate the underlying mechanisms of action through probing with small molecules.

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Agilent technologies

Agilent provides xCELLigence impedance-based, label-free, real time cell analysis system and NovoCyte flow cytometers.

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The RTCA DP Analyzer has three integrated stations for E-Plates 16 or CIM-Plates 16

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Located inside a tissue culture incubator, it is capable of switching any one of the wells on the E-Plate 96 to the RTCA Analyzer for impedance measurement

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Located inside a tissue culture incubator, it is capable of switching any one of the wells on any of six E-Plates to the RTCA Analyzer for impedance measurement

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Instrument can be integrated with automated liquid handling platforms or a robotic plate handler from different vendors

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