Cryopreservation Using Cross-Linked Bioactive Hydrogel Matrix Particles
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Solution Overview
Problem
Current cryopreservation methods for cells, particularly adherent cell cultures, often result in cell disruption during the detachment process, leading to damaged cells and the need for costly and time-consuming re-culturing post-thawing to restore cell integrity and viability.
Innovation Solution
A cross-linked bioactive hydrogel matrix is used to retain cells during cryopreservation, providing a scaffold for attachment and storage, which can be directly used in vivo without the need for re-culturing, utilizing a particulate form that allows cells to adhere to or be encapsulated within its structure, thereby minimizing damage and facilitating immediate use after thawing.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of operation
If cells are detached from adherent culture using trypsinization or mechanical scraping, then cells can be suspended for cryopreservation, but cell integrity and surface protein structure are damaged
Solution Approach 1:
The patent introduces a hydrogel matrix as an intermediary carrier that allows cells to be detached from the culture substrate without direct mechanical or enzymatic damage. Cells are trapped within the hydrogel matrix during cryopreservation, eliminating the need for harsh detachment methods while maintaining cell integrity and surface protein structure.
Solution Approach 2:
The patent changes the physical state and environmental parameters of the cell suspension by incorporating cells into a hydrogel matrix with specific rheological properties. This allows cells to remain in a protected three-dimensional environment during freezing, avoiding the harmful effects of conventional suspension methods.
2Duration of action of stationary object
If cells are subjected to conventional cryopreservation protocols, then cells can be stored for later use, but cell disruption occurs during the detachment and freezing process
Solution Approach 1:
The hydrogel matrix serves as a protective cushioning medium that is prepared beforehand to receive and protect cells during cryopreservation. This cushioning effect prevents cell disruption during the freezing process, maintaining high cell viability and integrity throughout storage duration.
3Reliability
If cell disruption is avoided during cryopreservation, then cell integrity is maintained, but conventional methods require complex detachment and re-culturing steps
Solution Approach 1:
The patent merges multiple functions into a single integrated system: the hydrogel matrix simultaneously serves as the culture substrate, the cryopreservation carrier, and the delivery vehicle. This consolidation eliminates separate detachment and re-culturing steps, simplifying the overall process while maintaining cell integrity.
Solution Approach 2:
The hydrogel matrix performs multiple functions: it provides the culture substrate for cell growth, serves as the cryopreservation matrix to protect cells during freezing, and acts as the delivery vehicle for cell transplantation. This multi-functionality reduces process complexity by eliminating the need for separate operational steps.
4Productivity
If adherent cells are placed in suspension for cryopreservation, then cells can be frozen, but cell surface proteins and membrane markers are destroyed or disturbed
Solution Approach 1:
The hydrogel matrix acts as a protective intermediary that encapsulates cells during cryopreservation, shielding cell surface proteins and membrane markers from mechanical and enzymatic damage. This allows efficient cryopreservation while preserving surface protein integrity for subsequent functional use.
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 integrity by reducing damage during the cryopreservation process, eliminating the need for re-culturing and allowing for direct in vivo application of cryopreserved cells, thus simplifying the cryopreservation process and improving cell recovery post-thawing.
Implementation Method 1
A cross-linked bioactive hydrogel matrix is used to retain cells during cryopreservation, providing a scaffold for attachment and storage
Implementation Method 2
cells should be cooled at a rate of around -1°C to -3°C per minute and thawed quickly by incubation in a 37°C waterbath
Data Source
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AI summary
The present invention is directed to methods of cryopreserving cells and cryopreserved cells prepared according to the methods. In specific embodiments, the method comprises combining cells with a cross-linked hydrogel matrix in particulate form, the matrix comprising a polyglycan cross-linked to a polypeptide and subjecting the combination to cryopreservation conditions. In further embodiments, the invention provides cell-seeded compositions comprising cells and a cross-linked bioactive hydrogel matrix in particulate form, the matrix comprising a polyglycan cross-linked to a polypeptide, wherein the composition has been subjected to cryopreservation conditions. The cryopreserved cells can be thawed and used in methods of treatment without the need for intervening steps to make the cells viable for in vivo use.