Assaying Adherent Cells Using High-Throughput Screening (HTS) by Cytometry

Traditionally, many cell-based flow cytometric assays have been limited to use with suspension cells, without being optimized for use with adherent cells. This is primarily due to the extra step required to lift and dissociate adherent cells to generate a single cell suspension suitable for flow cytometry. Neglecting adherent cells in flow analysis greatly restricts the potential application of many of these assays, since a large number of therapeutic drugs target receptors found on adherent cells particularly in the immuno-oncology area, such as treatments for breast or lung cancers. Therefore in order to achieve the maximum output from flow cytometry, it is essential that during  assay development we include the capabilities to interrogate adherent cell types, while avoiding shortfalls of conventional techniques that often:

• Provide bulk measurements (e.g. imaging) without a deeper investigation into effects on cell marker expression or cytokine release
• Require multiple workflows for quantification of different parameters, often necessitating multiple different instruments
• Lack physiological relevance towards in vivo conditions
• Involve lengthy, time-consuming workflows; requiring steps such as protocol optimization, fixation and repetitive washes

Here we present a portfolio of adherent cell assays for the iQue^® High-Throughput Screening (HTS) Cytometry Platform with easy-to-follow protocols for cell lifting and dissociation alongside validated reagent kits and analysis templates. We describe a range of assays across multiple research areas, including analysis of immune cell phenotype and function following killing of adherent tumor cells (in both 2D and 3D models); characterization of drug mechanism of action (MoA) in the context of ADCP and NK ADCC assays and assessment of therapeutic antibody internalization into adherent cell types. 

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