An in Vitro Microfluidic Platform Mimicking 3D Unidirectional Neural Circuits-based 3D Hydrogel Patterning for Brain Pathology

Abstract

Formation of unidirectional connection has limitations such as addition process, and growth of neurites simultaneously. So, it is necessary for an in vitro 3D unidirectional connectivity model that can simply mimic unidirectional neural circuits in brain. We developed a 3D in vitro unidirectional neural connection using a 5-channel microfluidic chip with patterning Matrigel. The microfluidic chip enables selective Matrigel patterning, unidirectional connection, and co-culture system by utilizing the micropillar structures. We confirmed separation between neurites and somas by Matrigel patterning. We achieved the separation after seeding cell-laden Matrigel in to channel 1 and 4. In addition, we optimized conditions for neuronal culture in respective channel, and coculture of two neuron groups. We also formed a unidirectional neural connection by different times between the axon from channel 2 to 3 and the dendrite from channel 4 to 1. One of important neuronal characteristics is the synapse, we confirmed the presynaptic neurons through synapsin 1 of immuno-fluorescence, and the synapses were mainly formed in channel 3. My approaches expected that it would be possible to identify the cause of mental disorders and develop fundamental treatments related to meso-cortical circuits in dopaminergic pathway.