Gas-Permeable Microplate Wells for Cell Cultivation
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for culturing cells in three-dimensional environments lack efficient systems for supporting cell aggregates and facilitating media exchange without disturbing the cellular environment, and existing microplates do not provide adequate oxygen supply and visualization capabilities.
Innovation Solution
A microplate design featuring gas-permeable and optically transparent wells with a support ledge and a clamping frame, along with a mold insert tool, to create microwell structures that support hydrogel and allow for gravitational media exchange and enhanced oxygen supply while enabling visualization of cell cultures.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If a traditional microplate design is used, then the device complexity is low, but the oxygen supply to cell aggregates is insufficient
Solution Approach 1:
The patent employs a gas-permeable membrane as the bottom surface of the well, which allows oxygen and other gases to diffuse through it. This porous/gas-permeable structure enables direct oxygen supply to the cell aggregates from the underlying gas phase, resolving the contradiction between maintaining simple device structure and improving oxygen availability.
2Reliability
If the well bottom is made gas-permeable, then oxygen supply is enhanced, but the structural stability of the well may be compromised
Solution Approach 1:
The gas-permeable membrane is implemented as a thin film structure that forms the bottom of the well. This thin film approach allows gas permeability while maintaining sufficient mechanical strength through proper material selection and membrane tensioning, resolving the contradiction between gas permeability and structural stability.
3Reliability
If the well bottom is made gas-permeable, then oxygen supply is enhanced, but the manufacturing precision becomes more difficult to achieve
Solution Approach 1:
The gas-permeable membrane can be manufactured using established techniques for creating porous or gas-permeable materials, such as track-etched membranes, anodized aluminum oxide, or sintered materials. These are well-established manufacturing processes that can achieve consistent pore sizes and gas permeability properties, resolving the manufacturing precision challenge.
4Reliability
If a support ledge is added to support hydrogel, then the support capability for cell aggregates is improved, but the device complexity increases
Solution Approach 1:
The well structure is segmented into distinct functional zones: the support ledge region for mechanical support of hydrogel and cell aggregates, and the gas-permeable membrane region for oxygen supply. This segmentation allows each region to perform its specific function optimally while maintaining overall structural integrity.
5Illumination intensity
If the gas-permeable sheet is made optically transparent, then visualization capability is improved, but the gas permeability may be reduced
Solution Approach 1:
The gas-permeable membrane is selected or engineered to be optically transparent while maintaining its gas permeability properties. Materials such as certain polymers with controlled pore structures can simultaneously provide optical clarity for visualization and adequate gas flux, resolving this contradiction.
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 microplate system effectively supports the growth and maintenance of cell aggregates by facilitating media exchange and oxygen supply, while allowing for the visualization of cellular structures, thereby improving the culturing and analysis of embryoid bodies and other multi-cellular bodies.
Implementation Method 1
a gas-permeable sheet secured to a lower portion of the array of wells, the gas-permeable sheet forming a bottom surface of at least a portion of each of the wells
Implementation Method 2
individual wells of the array of wells include a support ledge protruding from an interior surface of at least one well wall... the support ledge supporting the hydrogel within the individual wells
Implementation Method 3
the individual ones of the collars are coupled to corresponding ones of the wells via a friction fit
Data Source
AI summary
Disclosed are various embodiments for growing, culturing, monitoring, and analyzing embryoid bodies, fused embryoid bodies, spheroids, organoids, or other multi-cellular bodies in a microwell structure formed in one or more wells of an assay and culturing microplate. Hydrogel deposited into a well of the microplate and supported by a support ledge and the bottom surface of the well is molded into a microwell structure using a mold insert tool. In some examples, channels can be formed in the bottom of the microwell structure to allow for an exchange of fluid between a primary well section and a secondary well section of the well. The bottom surface of the assay and culturing microplate is optically transparent and gas-permeable.