Abstract
Microglia are the main inflammatory cell of the brain and have been implicated in the development and progression of several neuroinflammatory and neurodegenerative conditions such as Alzheimer’s disease (AD), Frontotemporal Dementia (FTD) and Amyotrophic Lateral Sclerosis (ALS). iPSC-derived microglia provide a scalable, reproducible and physiological relevant model with which to study this role. Axol Bioscience has developed a robust protocol to generate microglia, with over 30 lines successfully differentiated, including those derived from healthy and patient donors and gene-edited lines.
Microglia are highly plastic cells influenced by environmental cues to drive both pro- and antiinflammatory responses. As such, any dysregulation in their activation status can drive an exaggerated and sustained state of inflammation. axoCells iPSC-derived microglia provide a reliable platform to study phenotypes of healthy and diseased microglia and assess the ability of novel compounds to decrease inflammation. In this study, Axol used a range of assays to investigate the activation state of microglia in response to different stimuli, as well as assessing inter batch and inter assay variability of our control (ax0664) microglia.
HTRF assays for IL8, IL6 and TNFα release demonstrated consistent effects across batches of microglia, when stimulated with a concentration range of lipopolysaccharide (LPS). In addition, HTRF and ELISAs were used to assess activation states of microglia induced by IL-4. Further characterization by flow cytometry, investigated the effect of different stimuli and incubation times on CD80, CD83 and CD206 receptor expression. Additional profiling of ax0664 investigated the release of a broad range of cytokines, using O-link technology. Furthermore, TempO-Seq analysis was used to investigate transcriptomic profiles driven by different stimuli. Overall, this supports the ability of Axol’s human iPSC-derived microglia to polarise towards different activation states. axoCells iPSC-derived microglia can be used as a relevant platform for human drug discovery where they can help predict the action of new drugs and identify mechanisms involved in neuroinflammation.
