Immuno-oncology

Immune cell killing assays

WHY USE A KINETIC ASSAY?

Cell-based assays enable rapid evaluation of human biology in vitro, with substantial advantages in throughput and cost over tissue or animal models. However, many cell-based assays are endpoint assays, limited to a single snapshot in time. Axion BioSystems' Maestro Z platform now offers impedance-based cell analysis for real-time, continuous, label-free monitoring of your cells. Continuous data reveals the kinetics of cell-cell interactions and cell-drug responses for better mechanistic understanding without the time- and cost-intensive process of repeating multiple endpoint assays.

THE MAESTRO Ƶ ADVANTAGE

High sensitivity – want to evaluate killing kinetics with physiologically relevant effector cell to target cell ratios? The Maestro Z system is capable of evaluating low effector cell to target cell ratios that are more physiologically relevant to your research.

Continuous cell monitoring – the Maestro Z system performs noninvasive electrical measurements from the cultured cell population, circumventing the use of dyes/reporters that can perturb your cell model and confound results. Track activity over hours and weeks from the same population of cells label-free and in real-time.

Precise environment – there is no need for an additional cell culture incubator, saving valuable lab space and money. Maestro Z’s smart environmental chamber finely controls heat and CO₂ while rejecting electrical noise and mechanical vibrations.

Probe cell models in the same plate they were cultured in – other higher throughput platforms (e.g. flow cytometry) often require cell samples to be transferred into a single-cell suspension before testing. In the case of adherent cells this is not ideal since they exist as a functional network of interlinked cells. In addition, the cell harvesting process requires numerous handling steps. The Maestro Z system captures real-time cell data while preserving the morphological complexity of your cell model.

It's easy – just culture your cells in a CytoView-Z plate, load your plate into the Maestro Z system, and record your cell data. Axion's data analysis tools will do the rest. The AxIS Z software can even generate the publication-ready graphs you need.

HOW IT WORKS

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HOW IT WORKS

Learn how impedance assays work and the innovation behind Axion's products.

Axion Maestro Z impedance assay workflow

OVERVIEW

Getting started with Maestro Z couldn't be easier. Culture your cells in an Axion multiwell CytoView-Z plate [A]. Load this plate into the Maestro Z system and allow the environmental chamber to automatically equilibrate [B]. Track immune cell-mediated killing label-free and in real-time with AxIS Z software [C].

Axion Maestro Z impedance assay immuno-oncology immune cell kill assay recordings label-free

WHAT IS IMPEDANCE?

Axion's 96-well CytoView-Z plate has a recording electrode embedded in the culture surface of each well [D]. The Maestro Z platform uses impedance measurements (ohms, Ω) to quantify the presence of cells on the electrode [E]. To measure impedance, small electrical signals are delivered to the electrodes. Cell attachment, spreading, and cell-cell connections block these electrical signals and are detected as an increase in impedance. Since impedance is non-invasive and label-free, impedance assays can be used to quantify dynamic cellular responses over minutes, hours, and days [F].

Axion Maestro Z impedance electrode recordings cell proliferation cytotoxicity assay

CELLULAR IMPEDANCE RECORDINGS

Impedance measures how much electrical signal (teal-colored arrows) is blocked by the electrode-cell interface [G]. When the electrode is uncovered, electrical signal easily passes (thick arrows) and the impedance is low. When cells cover the electrode, less electrical signal passes (blocked arrows) and impedance is high. When cells die or detach, the impedance decreases back towards baseline [H].

APPLICATIONS

Discover how impedance-based assays are being used for immuno-oncology applications.

CANCER IMMUNOTHERAPY

Monitor effector-mediated cytolysis of target cancer cell lines

Glioblastoma (GBM) is an aggressive form of brain cancer that has no effective treatments and a prognosis of only 12–15 months. Immune system effector T cells hold promise for future cancer therapy due to their high specificity and innate cytotoxicity. The Maestro Z impedance assay provides a sensitive, label-free, nondestructive method to continuously monitor cancer cell proliferation and immune cell-mediated cytotoxicity, providing a powerful means of assessing immune cell potency.

U87MG glioma cells were seeded into the CytoView-Z plate at three different densities (n=12) and impedance was continuously monitored on the Maestro Z. After 24 hours, activated human T-cells were added in a 10:1 ratio to each well (n=4 wells for each cell density). The addition of activated human T-cells resulted in a decrease of the impedance signal consistent with T-cell-mediated lysis of the U87 cells, while untreated wells continued increasing, as shown (left) for 25k U87. Percent cytolysis was tracked in real-time (middle), enabling computation of KT50 for each density (right). A higher density of T-cells led to faster killing of U87 cells.