Symmetry Breaking of Human Pluripotent Stem Cells (hPSCs) in Micropattern Generates a Polarized Spinal Cord‐Like Organoid (pSCO) with Dorsoventral Organization

Abstract

Axis formation and related spatial patterning are initiated by symmetry breaking during development. A geometrically confined culture of human pluripotent stem cells (hPSCs) mimics symmetry breaking and cell patterning. Using this, polarized spinal cord organoids (pSCOs) with a self-organized dorsoventral (DV) organization are generated. The application of caudalization signals promoted regionalized cell differentiation along the radial axis and protrusion morphogenesis in confined hPSC colonies. These detached colonies grew into extended spinal cord-like organoids, which established self-ordered DV patterning along the long axis through the spontaneous expression of polarized DV patterning morphogens. The proportions of dorsal/ventral domains in the pSCOs can be controlled by the changes in the initial size of micropatterns, which altered the ratio of center-edge cells in 2D. In mature pSCOs, highly synchronized neural activity is separately detected in the dorsal and ventral side, indicating functional as well as structural patterning established in the organoids. This study provides a simple and precisely controllable method to generate spatially ordered organoids for the understanding of the biological principles of cell patterning and axis formation during neural development.

Design of neural organoids engineered by mechanical forces

“…Remarkably, connected neural organoids through projecting axons (also called connectoids) in a specific microdevice ( Kirihara et al, 2019 , Osaki and Ikeuchi, 2021 ) or within a tube of gel ( Cullen et al, 2019 ) can generate more complex neural circuits. On the other hand, a complex technique such as creating a concentration gradient of specific morphogens within neural organoids has been successfully simulated to mimic patterned structures during brain development ( Cederquist et al, 2019 , Rifes et al, 2020 , Seo et al, 2023 ).…”

Advancements in 2D and 3D In Vitro Models for Studying Neuromuscular Diseases

“…Furthermore, organoids are expected to be applied in transplantation treatments, where personalized organoids can be generated using stem cells derived from the patient. Organoids have been developed for use in various human tissues, including the brain, gut, kidney, liver, retina, spinal cord, and pancreas [2][3][4][5][6][7][8][9]. However, modeling diseases in which multiple components affect each other, such as neuromuscular disorders (NMDs), remains a challenge.…”