EDUCATIONAL MATERIAL (NENS)

NENS Cluster videos on glia and inflammation

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Astrocytes and microglia in neuroinflammation 

Glia and inflammation in Amyotrophic lateral sclerosis. Astrogliosis. Microglial phenotypes. Interaction of microglia and astrocytes with neurons. BBB breach and the role of astrocytes. Infiltration of immune cells in brain tissue and interaction with glia. Humoral factors of inflammation and glia. Comparative glial cellular morphology obtained by high resolution microscopy. Oxidative stress and microglia – release of cytokines. 

Dynamical measurements from neuroglia in vitro 

Preparation of primary rat microglia (and/or BV-2 cell line) /astrocytes for experiment. Dye loading – Ca2+– or ROS-sensitive dyes. Videomicroscopy setup. Recording software environment. Dynamical measurements of Ca2+ or ROS – transients. Signal analysis. 

Here we show a protocol for recording the electrical activity organoids on MEA. The premise is the comparison of organoids with and without microglia (Fagerlund and Dougalis et al., 2021). We go through the main steps in slicing an organoid, preparing it for recording and doing baseline recording and pharmacological stimulations. 

Protocol description starts at 2:10

Video edited by: 

Anssi Pelkonen

Camera:

Ilkka Fagerlund

MEA experiment:

Mireia Gomez-Budia

Special thanks:

Neuroinflammation research group lead by Tarja Malm

Publication:

Fagerlund, I.; Dougalis, A.; Shakirzyanova, A.; Gómez-Budia, M.; Pelkonen, A.; Konttinen, H.; Ohtonen, S.; Fazaludeen, M.F.; Koskuvi, M.; Kuusisto, J.; Hernández, D.; Pebay, A.; Koistinaho, J.; Rauramaa, T.; Lehtonen, Š.; Korhonen, P.; Malm, T. Microglia-like Cells Promote Neuronal Functions in Cerebral Organoids. Cells 2022, 11, 124. https://doi.org/10.3390/cells11010124

Some images also from:

Pelkonen, A.; Pistono, C.; Klecki, P.; Gómez-Budia, M.; Dougalis, A.; Konttinen, H.; Stanová, I.; Fagerlund, I.; Leinonen, V.; Korhonen, P.; Malm, T. Functional Characterization of Human Pluripotent Stem Cell-Derived Models of the Brain with Microelectrode Arrays. Cells 2022, 11, 106. https://doi.org/10.3390/cells11010106

Sources otherwise indicated in the video

Modeling Neuroinflammation in human context 

This lecture covers current knowledge on human-based models of neuroinflammation. These include iPSC-derived 2D and 3D models, possible differences between the models and their suitability for functional studies.  

How to culture human iPSC-derived microglia

We show the detailed protocol and practical tips for differentiation of microglia-like cells from induced pluripotent stem cells. We will introduce methodology to carry out quality control analysis of microglia throughout the differentiation protocol using flow cytometry and how to carry out functional studies for phagocytosis and migration using Incucyte Live Imaging.