Poster abstract - AD/PD 2024

Abstract title: Development of chimeric human-rodent functional and high content in vitro neuroinflammatory models

Poster number: P1014 / #1038

Authors: Joana De Abreu Carvalho1, Christina Nodin2, Paul Karila2
Affiliations: 1Apollo Therapeutics, Research, Cambridge, United Kingdom, 2Cellectricon AB, Mölndal, Sweden

Aims: Neuron-glia interactions and neuroinflammation may be dysfunctional in neurodevelopmental and neurodegenerative disorders. Complex cell biology in vitro models for investigating such interactions for drug discovery are lacking. We therefore set out to establish high-capacity, high-quality in vitro assays using rodent primary cortical cultures, consisting of neurons and astrocytes with human iPSC-derived (hiPSC) microglia. Our novel assays will investigate neuronal function and cell health and are promising tools for the testing of molecules with the potential to modulate neuroinflammation.
Methods: Chimeric cultures of rodent primary cortical cultures and hiPSC microglia were generated in 384 well-format. After 10-14 days, cell cultures were treated with LPS at different concentrations and timepoints. 1-5 days after LPS addition, neuronal, astrocytic and microglial morphology and TNF-alpha release was quantified. Further, optical electrophysiology was used to investigate effects on neuronal network activity.
Results: The chimeric model resulted in well-integrated hiPSC microglia, with morphology resembling homeostatic microglia, as well as healthy neurons and astrocytes. Addition of LPS resulted in a clear shift of hiPSC microglia to an amoeboid morphology as quantified by HCA, an increased TNF-alpha secretion and an increased electric field stimulation-evoked response. Changes in microglia were aligned with an activated pro-inflammatory status.
Conclusions: The results indicate that it is possible to generate biologically relevant neuroinflammation models for drug discovery, where both morphological and functional phenotypes such as cytokine release and neuronal network activity can be investigated for the functional profiling of molecules developed for clinical use.