A neuron, microglia, and astrocyte triple coculture model to study Alzheimer disease

Luchena, C.; Zuazo-Ibarra, J.; Valero, J.; Matute, C.; Alberdi, E.; Capetillo-Zarate, E.

doi:10.1101/2021.12.13.472367

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bioRxiv · 10.1101/2021.12.13.472367

A neuron, microglia, and astrocyte triple coculture model to study Alzheimer disease

Luchena, C.·Zuazo-Ibarra, J.·Valero, J.·Matute, C.·Alberdi, E.·Capetillo-Zarate, E.

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Abstract

Glial cells are essential to understand Alzheimers disease (AD) progression, given their role in neuroinflammation and neurodegeneration. There is a need for reliable and easy to manipulate models that allow studying the mechanisms behind neuron and glia communication. Currently available models such as cocultures require complex methodologies and/or might not be affordable for all laboratories. With this in mind, we aimed to establish a straightforward in vitro setting with neurons and glial cells to study AD. We generated a triple co-culture with neurons, microglia and astrocytes. Immunofluorescence, western blot and ELISA techniques were used to characterize the effects of oligomeric A{beta} (oA{beta}) in this model. We found that, in the triple co-culture, microglia increased the expression of anti-inflammatory markers Arginase I and TGF-{beta}1, and reduced pro-inflammatory iNOS and IL-1{beta}, compared with microglia alone. Astrocytes reduced expression of pro-inflammatory A1 markers AMIGO2 and C3, and displayed a ramified morphology resembling physiological conditions. Lastly, neurons increased post-synaptic markers, and developed more and longer branches than in individual primary cultures. Addition of oA{beta} in the triple coculture reduced synaptic markers and increased microglial activation, which are hallmarks of AD. Consequently, we developed a reliable model, where cells better resemble physiological conditions: microglia are less inflammatory, astrocytes are less reactive and neurons display a more mature morphology than in individual primary cultures. Moreover, we were able to recapitulate A{beta}-induced synaptic loss and inflammation. This model emerges as a powerful tool to study neurodegeneration and inflammation in the context of AD and other neurodegenerative diseases. Table of content image O_FIG_DISPLAY_L [Figure 1] M_FIG_DISPLAY C_FIG_DISPLAY Main pointsO_LIIn our model, microglia and astrocytes are less reactive, and neurons have a more mature morphology than in primary cultures. C_LIO_LIoA{beta} reduced synaptic markers and increased microglial activation. C_LIO_LIThis triple co-culture is a reliable tool to study neurodegeneration and gliosis in vitro. C_LI

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Luchena, C., Zuazo-Ibarra, J., Valero, J., Matute, C., Alberdi, E., Capetillo-Zarate, E.. 2021-12-14. A neuron, microglia, and astrocyte triple coculture model to study Alzheimer disease. https://doi.org/10.1101/2021.12.13.472367

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