Development of in vitro 3D unidirectional cerebral region circuit analytic platform by controlling the growth rate of neurites

초록

In vitro 3D neural circuit models can be actively used to identify disease pathogenesis and therapeutic drug screening. Especially, the hydrogel‐based 3D neural circuit can align the hydrogel related to cell behavior and cell signaling and can simulate the 3D structure and environment of the body. Therefore, it is necessary to develop and functionally analyze an in vitro unidirectional neural circuit similar to the three‐dimensional neural circuit connection in the body. However, due to the randomly directional growth of neurites in the hydrogel, it is difficult to form a unidirectional connection in the body. In addition, 3D neural circuit fluorescence imaging using immunostaining is difficult to perform functional analysis due to the limitation of microscopic resolution. So, we developed the in vitro 3D unidirectional cerebral region circuits by physical dissection of the cerebral organoids and alignment of collagen. First, the physical stimulus is applied to the neurites firmly bound in the organoid, the bonds of the inner neurites are broken, and the regrowth of the neurites to the outside is promoted. Secondly, the three‐dimensional environment of the neural circuit was formed based on the collagen scaffold, and the alignment of collagen fibers through shear stress in the pathway included the growth direction of the neurites. Finally, in vitro 3D unidirectional cerebral region circuits were measured functional activity using the 3D neural probe. Therefore, in vitro 3D unidirectional cerebral region circuit analytic platform can be used to analyze the mechanism of pathogenesis and develop treatment methods for mental disorders.