Organoid Production Using Microcontainers and Hydrogel Lids
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Solution Overview
Problem
Current organoid culture methods require exogenous extracellular matrix scaffolds, which are costly, have lot-to-lot variability, and are not physiologically representative, limiting their widespread deployment, especially for anchorage-dependent cells like human mammary epithelial cells that undergo anoikis in scaffold-free conditions.
Innovation Solution
A method involving microcontainers with hydrogel walls and lids that allow cells to culture without exogenous extracellular matrix or at low concentrations, using a culturing medium with biological colloids to achieve the necessary density and environment for organoid formation, enabling the accumulation of endogenously secreted matrix for physiological cues.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If exogenous extracellular matrix scaffolds are used for organoid culture, then cell survival and organoid development are improved, but cost increases and lot-to-lot variability occurs
Solution Approach 1:
The patent enables cells to produce their own extracellular matrix components endogenously, eliminating the need for exogenous matrix scaffolds. The culturing medium contains components that stimulate cells to secrete their own matrix proteins, allowing the system to be self-sufficient and avoiding the high costs and variability associated with commercial extracellular matrix products.
Solution Approach 2:
The invention extracts and removes the dependency on exogenous extracellular matrix scaffolds from the organoid culture system. By formulating a specialized culturing medium that promotes endogenous matrix production, the patent eliminates the need to add external matrix components, thereby removing the associated costs and quality control issues.
2Shape
If exogenous extracellular matrix scaffolds are used for organoid culture, then organoid structure formation is improved, but physiological representativeness deteriorates due to composition and structure discrepancies
Solution Approach 1:
The patent enables cells to produce their own extracellular matrix components endogenously, eliminating the need for exogenous matrix scaffolds. The culturing medium contains components that stimulate cells to secrete their own matrix proteins, allowing the system to be self-sufficient and avoiding the high costs and variability associated with commercial extracellular matrix products.
Solution Approach 2:
The invention changes the chemical composition parameters of the culturing medium to promote endogenous matrix production. By adjusting medium composition rather than adding exogenous matrix, the system achieves physiological relevance while maintaining proper organoid structure formation.
3Quantity of substance
If scaffold-free culture conditions are used for anchorage-dependent cells, then exogenous matrix requirements are eliminated, but cell viability deteriorates due to anoikis
Solution Approach 1:
The patent enables cells to produce their own extracellular matrix components endogenously, eliminating the need for exogenous matrix scaffolds. The culturing medium contains components that stimulate cells to secrete their own matrix proteins, allowing the system to be self-sufficient and avoiding the high costs and variability associated with commercial extracellular matrix products.
Solution Approach 2:
The invention introduces a specialized culturing medium as an intermediary that facilitates endogenous matrix production. This medium acts as a mediator between the scaffold-free environment and cell viability requirements, enabling cells to survive and thrive by producing their own matrix without needing exogenous scaffolds.
4Reliability
If high concentration of exogenous extracellular matrix is used, then cell survival is improved, but physiological relevance deteriorates due to discrepancies from bona fide basement membrane
Solution Approach 1:
The patent enables cells to produce their own extracellular matrix components endogenously, eliminating the need for exogenous matrix scaffolds. The culturing medium contains components that stimulate cells to secrete their own matrix proteins, allowing the system to be self-sufficient and avoiding the high costs and variability associated with commercial extracellular matrix products.
Solution Approach 2:
The invention changes the chemical composition parameters of the culturing medium to promote endogenous matrix production. By adjusting medium composition rather than adding exogenous matrix, the system achieves physiological relevance while maintaining proper organoid structure formation.
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
This approach allows for the production of organoids that exhibit contractility and physiological differentiation without the need for exogenous matrices, providing a more physiologically relevant and cost-effective method for organoid culture, with high throughput and scalability.
Implementation Method 1
the culturing medium has a higher density than the hydrogel lid. In certain embodiments, the culturing medium has a density between about 1.1 g/ml and about 1.2 g/ml and the hydrogel lid has a density of about 1.0 g/ml
Implementation Method 2
overlaying a hydrogel over the culture containing the cells such that the hydrogel forms a lid which is in direct contact with the surface of the culture to seal the culture
Data Source
AI summary
Disclosed are methods of producing organoids in the absence of any exogenous extracellular matrix or in the presence of an exogenous extracellular matrix at a concentration lower than gelling concentration, using microcontainers sealed with a hydrogel lid as well as organoids produced by this technique.