Microwell Array for Consistent 3D Cell Spheroid Culture
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Solution Overview
Problem
Current methods for culturing cells in three dimensions face challenges such as inconsistent spheroid size, shear-induced breakage, and inadequate cell density, leading to variations in assay results and inefficient gas exchange due to high media volumes.
Innovation Solution
The use of cell culture devices with microwells that have spheroid-inducing geometry and ultra-low binding coatings, along with capillary structures to prevent air trapping and facilitate liquid flow, allowing cells to self-assemble into consistent spheroids and maintain high density cultures.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If cells are grown in three dimensions as spheroids, then in vivo-like functionality is improved, but spheroid size consistency deteriorates
Solution Approach 1:
The invention divides the culture system into individual microwells, each acting as an independent compartment that confines cell growth. This segmentation prevents spheroids from merging and ensures uniform size by physically limiting the maximum diameter to the well diameter, while still allowing 3D growth and cell-cell interactions for in vivo-like functionality.
Solution Approach 2:
The invention changes the geometric parameter of the culture environment by using microwells with specific diameter dimensions (e.g., 200-500 μm). This parameter control directly determines spheroid size, ensuring consistency across cultures while maintaining the three-dimensional structure necessary for in vivo-like functionality.
2Productivity
If cell density is increased, then productivity is improved, but media volume requirements increase
Solution Approach 1:
By segmenting the culture into many small microwells, the system achieves high total cell density across the plate while each individual well requires minimal media volume. The cumulative effect of numerous low-volume wells provides high productivity without the need for large total media volumes.
Solution Approach 2:
The invention transitions from two-dimensional monolayer culture to three-dimensional spheroid culture within microwells. This dimensional change allows cells to grow vertically in 3D space rather than spreading horizontally, increasing cell density per unit volume of media and improving productivity without proportionally increasing media volume requirements.
3Productivity
If media height is increased to satisfy high density cell culture, then cell density is improved, but gas exchange rate deteriorates
Solution Approach 1:
The microwell structure segments the culture system such that each well has a controlled, limited height. This prevents excessive media depth that would hinder gas diffusion, while the high number of wells compensates for the smaller individual volume, maintaining high overall cell density without compromising gas exchange.
4Productivity
If conventional apparatuses are used for high density spheroid culture, then cell density can be increased, but spheroid size consistency and integrity deteriorate due to shear forces
Solution Approach 1:
The microwell array segments the culture environment into isolated compartments, allowing high cell density to be achieved across the plate without requiring mechanical agitation. Each well provides a quiescent environment that protects spheroids from shear forces, maintaining size consistency and integrity while achieving high overall productivity.
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 enables the formation of consistent, in vivo-like cell aggregates with improved gas exchange, maintaining cell health and productivity, and facilitating high-throughput research and drug testing.
Implementation Method 1
The microwells or wells may be non-adherent to cells or coated with an ultra-low binding material to make the wells non-adherent to cells
Implementation Method 2
the wells are structured and arranged to provide for the movement of liquid into and out of the wells without trapping air between the substrate and liquid or liquid droplets that are introduced into the wells
Data Source
AI summary
The present disclosure relates to apparatuses, systems and methods for culturing cells. In particular, devices and methods are provided for generation and culture of 3d cell aggregates.