Agitated Reactor Dissociation of Cell Aggregates
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Solution Overview
Problem
Current methods for dissociating cell aggregates in suspension cultures are inefficient and variable, particularly at larger scales, due to reliance on manual pipetting which is time-consuming, poorly controlled, and dependent on operator skill, limiting cell recovery and viability.
Innovation Solution
A method involving the use of an agitated reactor where cell aggregates are contacted with a dissociation reagent and exposed to a generated dissociation force, such as from a stirrer or forced fluid flow, to effectively dissociate the aggregates while maintaining high cell viability.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If manual pipetting is used to dissociate cell aggregates, then operator control is possible, but the process is time-consuming and highly variable
Solution Approach 1:
The patent replaces manual mechanical pipetting with an automated mechanical dissociation system that uses controlled shear force applied through a porous substrate. This substitution eliminates operator variability and time-consuming manual operations while maintaining consistent dissociation control through engineered mechanical parameters.
Solution Approach 2:
The invention changes the dissociation parameter from manual pipetting force to controlled shear force applied through a porous substrate. By defining shear force as a measurable and controllable parameter, the system achieves both precision and productivity, resolving the contradiction between control and efficiency.
2Quantity of substance
If manual dissociation is applied to large numbers of cells, then cell expansion is possible, but the method becomes ineffective
Solution Approach 1:
The patent segments the dissociation process into two functional components: a porous substrate that provides structural support and enables force distribution, and a dissociation reagent that chemically breaks cell-cell adhesion. This segmentation allows the system to handle large cell numbers effectively by distributing the dissociation force across the entire cell population simultaneously.
Solution Approach 2:
The porous substrate acts as an intermediary between the applied shear force and the cell aggregates. It distributes the mechanical force uniformly across the cell population, enabling effective dissociation of large numbers of cells that would be impossible to handle with manual pipetting.
3Productivity
If agitation force is increased to dissociate aggregates, then dissociation efficiency improves, but cell damage increases
Solution Approach 1:
The patent applies local quality by using a porous substrate with specific pore sizes and structures that localize and control the shear force application. The substrate's physical properties are optimized to provide sufficient dissociation force while protecting cells from excessive mechanical damage, resolving the contradiction between dissociation efficiency and cell viability.
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 method enables efficient dissociation of cell aggregates at larger scales with high cell viability (>90%) and reduces operator dependence, facilitating scalable cell culture and production.
Implementation Method 1
contacting the cell aggregates with a dissociation reagent
Implementation Method 2
generating a dissociation force in the agitated reactor; the dissociation force is generated by movement of a stirrer, impeller, paddle, or wheel
Data Source
AI summary
A method for dissociating cell aggregates in an agitated reactor. The method comprises providing a cell culture comprising cell aggregates in the agitated reactor, contacting the cell aggregates with a dissociation reagent, generating a dissociation force in the agitated reactor and exposing the contacted cell aggregates to the generated dissociation force under conditions sufficient to dissociate the contacted cell aggregates. The method may be used in a process for passaging cells and/or generating dissociated differentiated cells from stem and/or progenitor cells.