抄録
In vitro 3D and organoid culture methods that emulate the intricate complexity of cell populations and extracellular matrix (ECM) components In vitro 3D significantly contribute to advancing our understanding of various biological phenomena. However, achieving precise control over the complex shapes, architectures, and interactions among different tissues within cultured organoids remains a challenge. Current organoid development heavily relies on cellular self–organization, yet the uniform culture conditions In vitro 3D fall short of providing accurate spatial cues to cells. Conversely, leveraging engineering principles offers a promising avenue to customize the design, composition, and construction of organoids based on specific research objectives. We have developed an in vitro experimental platform for the organoid culture to design and control microenvironment. The simple cube device, which comprises a polycarbonate frame with rigid agarose walls and an inner ECM hydrogel, can be used as a carrier of organoid to (i) control the spatial distribution of cells by employing 3D–printed carbohydrate moulds to create cell seeding pockets in the ECM hydrogel, (ii) design tissues with localized ECM by isolating ECM hydrogels of varying the composition or stiffness in separate compartments, (iii) facilitate integration with microfluidics to generate the concentration gradient of morphogens to direct cell growth and differentiation, (iv) assemble multi–CUBE with organoids or tissues to express tissue-tissue interactions. By employing above technologies, we were able to replicate the notochord signal during the development of the neural tube. The Shh gradient, facilitated through a 100 μm slit on a CUBE device, was applied to the neural plate, resulting in the generation of a localized expression pattern on the neural tube organoid.
