Organoid Microwell Positioning for Accessible 3D Network Culture
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Solution Overview
Problem
Existing methods for culturing organoids in three-dimensional networks face challenges with reproducibility, accessibility, and efficient three-dimensional extension due to unreliable positioning and limited accessibility of cells.
Innovation Solution
A microwell system with a main body and positioning members that allows for fixed, reproducible positioning of organoids, keeping them at a distance from the wall and bottom, enabling standardized manipulation and measurement, and facilitating three-dimensional growth.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Adaptability or versatility
If cells are injected into a hydrogel to form an organoid, then three-dimensional culture is achieved, but reliable positioning and accessibility are compromised
Solution Approach 1:
The invention divides the culture system into distinct functional components: a microwell structure for positioning, a hydrogel layer for three-dimensional culture, and an electrode layer for measurement. This segmentation allows each component to fulfill its specific function optimally while working together as an integrated system.
Solution Approach 2:
The microwell acts as an intermediary structure between the rigid substrate and the soft hydrogel containing the organoid. It provides mechanical support and precise positioning without directly contacting or compressing the organoid, thereby maintaining both positioning precision and three-dimensional culture integrity.
2Ease of operation
If organoids are cultured at the bottom of a reservoir, then ease of access is improved, but three-dimensional extension is impaired
Solution Approach 1:
The invention transitions from two-dimensional bottom culture to three-dimensional suspension culture within the microwell. The organoid is positioned in the middle layer where it can extend freely in all three spatial dimensions while the microwell structure ensures it remains within the accessible culture volume.
3Ease of operation
If organoids are positioned close to the wall, then accessibility is improved, but three-dimensional growth is restricted
Solution Approach 1:
The microwell structure creates a localized positioning zone that provides precise spatial control without imposing constraints on the organoid's three-dimensional network structure. The positioning function is localized to the micrawell geometry while the organoid maintains its natural growth pattern within this defined space.
Data Source
Figure 1A~1B
Figure 1C~1E
Figure 2A~2B
AI summary
The present invention relates to a microwell for culturing an organoid at a distance from the wall of the microwell and allowing development of a three-dimensional network, and to a device for measuring electrical activity of and/or stimulating cells in a three-dimensional network.