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FCDI Poster Presentation
Title: Modelling Neuroinflammation using iPSC-derived Engineered Microglia
Authors: Beatriz Freitas, Michael McLachlan, Sarah J Dickerson, Christie A Munn, Sarah A Burton, Abbey Musinsky, Madelyn Goedland, Anne Strouse, Deepika Rajesh, Simon Hilcove and Eugenia Jones
Abstract: Neurodegenerative and neurodevelopmental disorders can display aspects of neuroinflammation with microglial cells as crucial players during this detrimental stage. Microglia, the resident immune cells of the central nervous system, are a necessary cell type for neuronal homeostasis and are also responsible for synaptic pruning and brain development. Human iPSC-derived microglia can serve as an authentic preclinical tool for understanding the pathobiology of neurodegenerative and neurodevelopmental diseases. The present study involves the generation and characterization of genome engineered iPSC-derived iCell(R) Microglia to facilitate disease modeling of neurodevelopmental (Rett Syndrome) and neurodegenerative disorders (Alzheimer’s and Parkinson’s Disease). iCell Microglia generated from both the engineered and non-engineered iPSC clones offer unique isogenic pairs for research applications.
Microglia were derived by first by differentiating iPSCs into purified hematopoietic progenitor cells (HPCs), which were then further differentiated into microglia using 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 were highly characterized by assessing: (i) morphology; (ii) protein expression of TREM2, P2RY12, CX3CR1, IBA1, CD33 and CD45 by flow cytometry; (iii) phagocytic function using pHrodo-labelled BioParticles and aggregated amyloid beta (iv) quantification of neuroinflammatory molecules by multiplex Luminex; (v) measurement of soluble TREM by ELISA; and finally (vi) RNAseq analysis. Cryopreserved microglia retained purity and function comparable to pre-cryopreserved end-stage microglia. These results identified critical differences between wild type (WT) and engineered microglia in survival, phagocytosis kinetics, and expression levels of molecules involved in neural inflammation. Thus,these data demonstrate that iPSC-derived isogenic WT and engineered microglia could serve as a powerful tool to gain insight into various physiological and pathological conditions associated with neurological disorders and serve as a reliable in vitro disease model for complex neuronal diseases. .