Stably Inverted Organoids for Direct Apical Surface Access
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Solution Overview
Problem
Existing 3D tissue organoid systems suffer from non-uniformity in size and shape, difficulty in accessing the interior surface for studying cancer, bacterial, or viral invasion, and inconvenient handling, particularly due to their spherical or non-spherical structures with hollow or partially cell-filled interiors and apical-in polarity.
Innovation Solution
The development of stably-inverted organoids with a hollow lumen and epithelial cells exposed externally, allowing easier access for invasion studies, featuring a tissue layer with a basement membrane and extracellular matrix mixture, and comprising various cell types such as epithelial and stromal cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If conventional apical-in organoid structures are used, then the basement membrane forms around the basal side providing structural integrity, but the interior lumen becomes difficult to access for studying cancer, bacterial, or viral invasion
Solution Approach 1:
The patent inverts the traditional apical-in organoid structure to create an apical-out configuration where the apical surface of epithelial cells faces the exterior environment. This inversion is achieved by culturing organoids in a way that reverses their polarity, allowing the apical surface to be directly accessible from the outside without requiring microinjections into the lumen. The basement membrane remains intact but is repositioned relative to the apical surface, enabling direct access to epithelial cells for studying invasion while maintaining structural integrity.
2Productivity
If self-organized organoids are cultured, then they provide scalable platforms for high-throughput drug screenings, but they exhibit non-uniformity in size, shape, and cellular composition
Solution Approach 1:
The patent employs parameter changes in the culture conditions to achieve uniform organoid formation. Specific parameters including extracellular matrix composition (Matrigel concentration), growth factors (EGF, HB-EGF concentrations), and culture duration are optimized to produce organoids with consistent size, shape, and cellular composition. The method establishes controlled culture parameters that promote uniform spheroid formation and maturation, enabling reproducible high-throughput screenings while maintaining scalability.
3Ease of operation
If microinjections are used to access the lumen of micrometer-sized organoids, then cancer cells of interest can be introduced, but the procedure becomes time-consuming and technically demanding
Solution Approach 1:
The patent inverts the organoid structure so that the apical surface faces outward, eliminating the need for microinjections. Cancer cells or other test materials can be directly applied to the exterior apical surface, allowing rapid introduction without time-consuming microinjection procedures. This inversion fundamentally changes the access method from internal lumen injection to external surface application, dramatically reducing procedural time and technical complexity.
Data Source
AI summary
An exemplary embodiment of the present disclosure provides a 3D structure comprising a tissue layer having a first surface defining an interior chamber and an opposing second surface supporting a first plurality of cells outwardly positioned from the interior chamber, wherein the interior chamber comprises an extracellular matrix mixture. The present disclosure also provides a method of making a 3D structure, the method comprising mixing an extracellular matrix mixture at a first temperature with a culture medium at a second temperature, the second temperature greater than the first temperature, culturing a first plurality of cells in the extracellular matrix mixture and culture medium, and forming a 3D structure having an interior chamber enclosed by the first plurality of cells configured to interface with an environment external to the 3D structure.