Microtissue Propagation Device with Segmented Well and Bottleneck
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Solution Overview
Problem
Existing devices for propagating microtissues in two-dimensional cell cultures face challenges in preventing cell adhesion to the culture vessel, leading to potential aspiration of microtissues during medium changes, especially in automated handling procedures, which can disrupt the three-dimensional aggregation of cells.
Innovation Solution
A device with a well configuration featuring an open upper section for bulk culture medium handling and a lower section with a smaller cross-sectional area for microtissue formation, where the lower section is eccentrically arranged and has an ultra-low adhesion surface, along with a pipetting stage and chute to facilitate fluid handling without disturbing the microtissue, and a multiwell plate design that prevents fluid spreading and supports microscopic inspection.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Stability of the object's composition
If non-adhesively coated multiwell plates are used for propagating microtissues, then cell adhesion to the culture vessel is prevented and three-dimensional aggregation is promoted, but the microtissues become vulnerable to inadvertent aspiration along with the culture medium during medium changes
Solution Approach 1:
The well is divided into two distinct sections: an open upper section for bulk culture medium handling and a lower section with a bottleneck configuration for microtissue formation. This segmentation allows the microtissue to be confined in the lower section while medium is exchanged in the upper section, preventing aspiration of the microtissue during medium changes while maintaining its three-dimensional aggregation.
Solution Approach 2:
The bottleneck configuration acts as an intermediary structure between the open upper section and the lower section. It serves as a physical barrier that prevents the microtissue from being aspirated into the upper section during medium exchange, while still allowing the microtissue to maintain its three-dimensional aggregation in the lower section.
2Adaptability or versatility
If non-adhesively coated multiwell plates are used to prevent cell adhesion, then microtissue formation is enabled, but automatic handling procedures become problematic due to the vulnerability of microtissues to aspiration
Solution Approach 1:
By segmenting the well into an open upper section and a lower section with a bottleneck, the device enables automatic handling procedures. The bottleneck confines the microtissue in the lower section while allowing automatic medium exchange systems to operate in the upper section without risking microtissue aspiration, thus enabling automation while maintaining microtissue formation capability.
Solution Approach 2:
The bottleneck configuration serves as a protective intermediary that allows automatic handling systems to exchange medium in the open upper section without directly interacting with the microtissue in the lower section. This enables automatic handling procedures to be implemented safely while preserving the microtissue formation capability.
3Ease of operation
If conventional two-dimensional cell culture flasks are used, then cell propagation is simplified, but cell adhesion to the inner lining impairs three-dimensional aggregation required for microtissue formation
Solution Approach 1:
The well is segmented into an open upper section that allows simplified medium handling similar to conventional flasks, and a lower section with a bottleneck that promotes three-dimensional aggregation. This segmentation maintains ease of operation for medium exchange while enabling the three-dimensional cell aggregation necessary for microtissue formation.
Solution Approach 2:
Different sections of the well have different functional qualities: the open upper section provides ease of operation for medium handling, while the lower section with the bottleneck provides the specific local condition needed for three-dimensional aggregation. This local differentiation resolves the contradiction between operational simplicity and three-dimensional 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
The device effectively reduces the risk of microtissue aspiration during medium changes, allows for automatic handling, and enhances microscopic inspection by maintaining the microtissue's integrity and facilitating its formation and analysis.
Implementation Method 1
the lower section of said at least one well is provided with an ultra-low adhesion surface
Data Source
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Figure 3a~3c
AI summary
Disclosed are devices for propagating at least one microtissue, methods for manufacturing such devices, and the use us such devices, said devices comprise at least one well, wherein said at least one well comprises an open upper section comprising an open upper end and an open lower end, and a lower section comprising an open upper end and a bottom end, wherein the upper section and the lower section are in fluid communication, and wherein the bottom end of the lower section of at least one well of said plurality of wells is provided with a well bottom, said lower section of said at least one well has a smaller cross-sectional area than the open upper section of the same well.