Developing a perfusable 3D brain model in a multi-well chip system based on the in-vitro differentiation of the iPSC- derived radial glia cells

Authors

• S Anmalsetty
• SJ Silva
• G Vitale
• AJ Walke
• DA Scheiblin
• LA Ridnour
• DA Wink
• SJ Lockett
• WF Heinz

Abstract

Among the various 3D tissue/organ model developed in the past decade, the 3D brain model is one of the most challenging tissue models to engineer because of the physiological complexity and long maturation time for neural cells to be functional and the unique blood-characteristics of the blood-brain-barrier (BBB). Neural organoids are stem cell derived complex tissues models used to study brain development, but most lack of vascular system and immune cells. Based on our previous work developing a bioprinted 3D neurovasculature tissue model, we identified the Notch and Wnt signaling pathways as active during neurovascular formation. We hypothesized that activation of these pathways involved in neurogenesis would create an in-situ environment permissive of neuronal differentiation within a perfusable vascular network. We use iPSC-derived radial glia cells, which can differentiate into neuronal cells, astrocytes, and oligodendrocytes, and mixed them with primary human microvascular endothelial cell, primary human astrocyte, and primary human pericytes, in a fibrinogen-based hydrogel of a microfluidics channel of a 64-chips Mimetas Organograft plate. A perfusable microvascular network formed after 5 days of culture, and TUJ1 positive neuronal cells differentiated from iPSC radial glia cells were observed after two weeks. Weak signal for mature neuronal maturation cell marker, MAP2, and oligodendrocyte cell marker, Oligo2, was detected at day 21, indicating maturation of a neural tissue model. Functional assays on vascular permeability and neural activity are being conducted to demonstrate high physiological relevance of this tissue model and the potential application in biomedical/pharmacological studies.

Scientific Focus Area: Biomedical Engineering and Biophysics

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