Cell Aggregate Dissociation Using Metal Ion Adsorbents
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Solution Overview
Problem
Existing methods for dissociating cell aggregates in cell culture, particularly for pluripotent stem cells, are complex and cause significant damage to the cells, necessitating multiple steps and centrifugation for removing dissociation agents.
Innovation Solution
Using a water-insoluble metal ion adsorbent, such as a weakly acidic cation exchange resin, to dissociate cell aggregates without the need for centrifugation, allowing for easier and less damaging passage of cells.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If a low molecular weight chelating agent or enzyme is used as a dissociation agent, then cell aggregates can be dissociated into single cells, but the dissociation agent needs to be removed by centrifugation after the reaction
Solution Approach 1:
The patent extracts the harmful aspect (need for centrifugation) by using a water-soluble dissociation agent that can be easily removed through simple filtration or decantation, eliminating the need for complex centrifugation steps while maintaining effective cell aggregate dissociation
Solution Approach 2:
The patent introduces a water-soluble dissociation agent as an intermediary substance that facilitates cell aggregate dissociation and can be subsequently removed through simple filtration or decantation, acting as a mediator between the cell aggregates and the desired single-cell state without requiring centrifugation
2Productivity
If physical dissociation by agitation force is used, then cell aggregates can be separated, but the process is complicated and time-consuming
Solution Approach 1:
The patent replaces mechanical agitation forces with a chemical dissociation approach using a water-soluble dissociation agent, substituting the mechanical system (agitation reactor) with a chemical system that achieves dissociation through chemical action followed by simple physical removal of the agent
3Reliability
If multiple steps of separation, washing, and resuspension are performed, then cells can be passed, but the process becomes extremely complicated
Solution Approach 1:
The patent merges multiple separate steps (separation, washing, resuspension) into a streamlined process where the water-soluble dissociation agent is added to dissociate aggregates, then simply filtered or decanted to remove the agent, combining multiple operations into a more efficient single procedure
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
Efficient dissociation of cell aggregates with reduced cell damage, enabling large-scale production of pluripotent stem cells for research and medical applications.
Implementation Method 1
subjecting the cell aggregate to a treatment with a water-insoluble metal ion adsorbent
Implementation Method 2
the metal ion adsorbent has sedimentation property
Data Source
AI summary
The present invention aims to provide a method for dissociating cell aggregates, which are formed in cell culture, particularly in the cell passaging step, more easily and with less damage to cells, a method for culturing cells using said method, and a cell aggregate dissociation agent therefor. The present invention relates to a method for dissociating cell aggregates, including subjecting cell aggregates to a treatment with a water-insoluble metal ion adsorbent.