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Microglia are immune cells resident in the central nervous system (CNS) responsible for fundamental physiological and pathological processes. Microglia support neuronal homeostasis and facilitate neuronal network formation by synaptic pruning. Surveillant microglia can activate, adapt and respond accordingly to specific stimulus. Microglia dysfunction has been implicated in neurodegeneration afflictions, including Parkinson’s and Alzheimer’s diseases, and neurodevelopmental disorders, such as Rett syndrome. The limited availability and inconsistency of primary human microglia has constrained research and therapeutic progress.
iCell® Microglia are based on technology developed by the Blurton-Jones laboratory (Abud et al. Neuron 2017.) for which FujiFilm Cellular Dynamics Inc. has an exclusive license from the University of California-Irvine. iCell Microglia overcome the limitations of primary microglial cells by offering a consistent, high-quality microglia derived from human induced pluripotent stem cells, enabling the study of cytokine signaling, synaptic transmission, and plasticity in normal CNS function and during disease progression.
Using the IncuCyte® Live-Cell Analysis System (Sartorius AG), Staphylococcus aureus (SA) BioParticles® Conjugate (ThermoFisher Scientific) were added to iCell Microglia cultures to assess the kinetics of phagocytosis
iCell Microglia were thawed into 96-well plates and allowed to recover for three days prior to treatment (using lipopolysaccharide (100ng/mL) or IFN gamma (50ng/mL). After 24 hr, cytokine levels in the supernatant were assayed using the Luminex Multiplex Assay. Specific cytokine signatures were observed for each stimulus.
Based on published data describing gene expression and signatures of several microglial stages (Abud et al, 2017a>), iCell Microglia represent a valid in vitro model for CNS research. These cells express TREM2, CXCR4, P2RY12, IBA1, and PU.1 microglial markers at over 80% in the overall cell population. Physiologically, iCell Microglia demonstrate critical responses to stimuli including signature cytokine release, and the phagocytosis of bioparticles. Flow cytometry analysis of surface markers shows that iCell Microglia exhibit a characteristic microglia signature.
iCell Microglia are semi-adherent cells amenable to a variety of applications. iCell Microglia express key transcription factors and markers (CX3CR1, IBA1, TMEM119 and P2RY12) that are characteristically found in adult human microglia and are able to phagocytose and secret cytokines upon stimulation. Cell-based Assays Immunofluorescence Phagocytosis Amyloid-beta pHrodo™ Red labeling Figure 1: iCell Microglia Physiology (A) iCell Microglia secrete specific cytokines upon stimulation. (B) iCell Microglia phagocytose BioParticles® in a concentration-dependent manner. Advantages