A 3D human co-culture to model neuron-astrocyte interactions in tauopathies

Biological Procedures Online

Methodology

Paulien Cornelissen-Steijger¹  Vivi M. Heine¹  Leo S. Price²  Claudia Sestito³  Kevin L. Batenburg⁴  Jan R. T. van Weering⁵  Wiep Scheper⁵ 

[1] Amsterdam UMC location Vrije Universiteit Amsterdam, Department of Child and Adolescent Psychiatry, Amsterdam Neuroscience, De Boelelaan 1085, 1081 HV, Amsterdam, The Netherlands;Department of Complex Trait Genetics, Center for Neurogenomics and Cognitive Research, Vrije Universiteit Amsterdam, Amsterdam Neuroscience, De Boelelaan 1085, 1081 HV, Amsterdam, The Netherlands;Crown Bioscience Netherlands B.V. (Formerly OcellO B.V.), Leiden, The Netherlands;Crown Bioscience Netherlands B.V. (Formerly OcellO B.V.), Leiden, The Netherlands;Amsterdam UMC location Vrije Universiteit Amsterdam, Department of Child and Adolescent Psychiatry, Amsterdam Neuroscience, De Boelelaan 1085, 1081 HV, Amsterdam, The Netherlands;Department of Functional Genomics, Center for Neurogenomics and Cognitive Research, Vrije Universiteit Amsterdam, Amsterdam Neuroscience - Neurodegeneration, De Boelelaan 1085, 1081 HV, Amsterdam, The Netherlands;Department of Functional Genomics, Center for Neurogenomics and Cognitive Research, Vrije Universiteit Amsterdam, Amsterdam Neuroscience - Neurodegeneration, De Boelelaan 1085, 1081 HV, Amsterdam, The Netherlands;Amsterdam UMC location Vrije Universiteit Amsterdam, Department of Human Genetics, Amsterdam Neuroscience - Neurodegeneration, De Boelelaan 1085, 1081 HV, Amsterdam, The Netherlands;

关键词: hiPSC-derived neurons;    Astrocytes;    Neuron-astrocyte interaction;    3D co-culture;    Tau aggregation;   

DOI  :  10.1186/s12575-023-00194-2

 received in 2022-09-02, accepted in 2023-02-17,  发布年份 2023

来源: Springer

【 摘 要 】

BackgroundIntraneuronal tau aggregation is the major pathological hallmark of neurodegenerative tauopathies. It is now generally acknowledged that tau aggregation also affects astrocytes in a cell non-autonomous manner. However, mechanisms involved are unclear, partly because of the lack of models that reflect the situation in the human tauopathy brain. To accurately model neuron-astrocyte interaction in tauopathies, there is a need for a model that contains both human neurons and human astrocytes, intraneuronal tau pathology and mimics the three-dimensional architecture of the brain.ResultsHere we established a novel 100–200 µm thick 3D human neuron/astrocyte co-culture model of tau pathology, comprising homogenous populations of hiPSC-derived neurons and primary human astrocytes in microwell format. Using confocal, electron and live microscopy, we validate the procedures by showing that neurons in the 3D co-culture form pre- and postsynapses and display spontaneous calcium transients within 4 weeks. Astrocytes in the 3D co-culture display bipolar and stellate morphologies with extensive processes that ensheath neuronal somas, spatially align with axons and dendrites and can be found perisynaptically. The complex morphology of astrocytes and the interaction with neurons in the 3D co-culture mirrors that in the human brain, indicating the model’s potential to study physiological and pathological neuron-astrocyte interaction in vitro. Finally, we successfully implemented a methodology to introduce seed-independent intraneuronal tau aggregation in the 3D co-culture, enabling study of neuron-astrocyte interaction in early tau pathogenesis.ConclusionsAltogether, these data provide proof-of-concept for the utility of this rapid, miniaturized, and standardized 3D model for cell type-specific manipulations, such as the intraneuronal pathology that is associated with neurodegenerative disorders.

【 授权许可】

CC BY   
© The Author(s) 2023

【 图 表 】

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