2024 AIChE Annual Meeting
Determining the Effects of Geometric Confinement on the Fusion of Human Embryoid Bodies
While human cerebral organoids(hCO) serve as effective 3-dimensional models for experimentation, they often lack reproducibility and precise spatial organization, limiting their utility in research and potential therapeutic applications. Prior literature has suggested that spatial and mechanical factors can influence aspects of hCO development. This study explores the impact of geometric confinement on the fusion and development of embryoid bodies (EBs), to potentially enhance control over the reproducibility and spatial organization of the hCOs they differentiate into. To explore the relationship between geometric confinement and EB fusion, we designed and 3D-printed custom macrowells with different shapes to confine and guide EB fusion. We created an agarose mold of these macrowells and seeded -EBs within them, then following a human cortical organoid protocol. EBs were allowed to develop and fuse within the macrowells for 24 days and notable differences were observed between different geometries and EB developmental stages. We have observed that some geometries worked better for EB fusion than the others. After developing for 30 days within the macrowells, the successfully fused EBs underwent fixation, cryosectioning, and immunostaining for markers such as MAP2, EMX1, and SOX2. We are currently in the process of analyzing the effects of geometric confinement on the organizational differences between these fused EBs using confocal microscopy. If the results suggest that geometric confinement does have an effect on the organizational differences within the fused EBs, researchers may be able to fuse organoids within specific shapes tailored to particular experimental needs or to more closely mimic specific organ structures. These advancements could significantly enhance the utility of human cerebral organoids as a model for studying human development, disease progression, and drug responses.