Cryoprotective Solution for Cardiomyocyte Aggregate Freezing
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Solution Overview
Problem
Current methods for freezing pluripotent stem cell-derived cardiomyocytes result in unstable and low cell viability due to damage from proteolytic enzymes and loss of three-dimensional structure, making it difficult to reproduce uniform aggregates and maintain electrophysiological functional patterns post-thawing.
Innovation Solution
A method involving immersion of cardiomyocyte aggregates in a cryoprotective solution followed by freezing, which helps preserve the structural integrity and viability of the cells, allowing for the maintenance of electrophysiological properties post-thawing.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If cardiomyocytes are dispersed with proteolytic enzymes and frozen in a single cell state, then freezing operation is simplified, but cell viability becomes unstable and low due to enzyme damage and loss of three-dimensional structure
Solution Approach 1:
The patent applies preliminary action by immersing the cardiomyocyte aggregates in a cryoprotective solution before freezing. This preliminary treatment protects the cells from freezing damage and maintains their viability, eliminating the need for proteolytic enzyme dispersion while ensuring stable and high cell viability after thawing.
2Device complexity
If cardiomyocytes are dispersed and frozen in a single cell state, then freezing process is simpler, but electrophysiological functional patterns are not reproduced after thawing due to loss of aggregate structure information
Solution Approach 1:
The patent changes the physical state parameter by maintaining the cardiomyocytes in an aggregate form rather than dispersing them into single cells. The cryoprotective solution enables freezing of intact aggregates, preserving their three-dimensional structure and intercellular binding information, which allows electrophysiological functional patterns to be reproduced after thawing.
3Ease of manufacture
If proteolytic enzymes are used to disperse aggregates, then cell separation is achieved, but uniform aggregate reproduction becomes difficult due to enzyme damage
Solution Approach 1:
The patent extracts and eliminates the use of proteolytic enzymes from the process. By directly freezing the cardiomyocyte aggregates in their natural state with cryoprotective solution, the method avoids enzyme-induced damage and enables consistent reproduction of uniform aggregates after thawing, while still achieving effective cell separation through the freezing process itself.
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 enhances cell viability and maintains electrophysiological properties, enabling the practical use of frozen cardiomyocytes in applications such as drug screening and transplantation with high reproducibility and stability.
Implementation Method 1
immersing an aggregate of pluripotent stem cell-derived cardiomyocytes in a cryoprotective solution
Implementation Method 2
freezing the aggregate immersed in the cryoprotective solution
Data Source
AI summary
The disclosure provides a method of freezing an aggregate of pluripotent stem cell-derived cardiomyocytes, comprising:(i) immersing an aggregate of pluripotent stem cell-derived cardiomyocytes in a cryoprotective solution; and(ii) freezing the aggregate immersed in the cryoprotective solution.The disclosure also provides a frozen aggregate of pluripotent stem cell-derived cardiomyocytes frozen by the method.