Microfibrous Scaffolds Guide Stem Cell Lumenogenesis and Brain Organoid Engineering.

3D organoids have been widely used as tractable in vitro models capable of elucidating aspects of human development and disease. However, the manual and low throughput culture methods coupled with a low reproducibility and geometric heterogeneity restricts the scope and application of organoid research. Combining expertise from stem cell biology and bioengineering offers a promising approach to address some of these limitations. Here, we use melt electrospinning writing to generate tuneable grid scaffolds that can guide the self-organization of pluripotent stem cells into patterned arrays of embryoid bodies. We show that grid geometry is a key determinant of stem cell self-organization, guiding the position and size of emerging lumens via curvature-controlled tissue growth. We report two distinct methods for culturing scaffold-grown embryoid bodies into either interconnected or spatially discrete cerebral organoids. These scaffolds provide a high-throughput method to generate, culture and analyse large numbers of organoids, substantially reducing the time investment and manual labour involved in conventional methods of organoid culture. We anticipate that this methodological development will open up new opportunities for guiding pluripotent stem cell culture, studying lumenogenesis, and generating large numbers of uniform organoids for high throughput screening. This article is protected by copyright. All rights reserved.