Collagen Cell Carrier Cryopreservation Mechanical Stability
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Solution Overview
Problem
Current methods for cryopreservation of cells and tissue cultures face challenges with cell vitality loss during freezing and thawing, and existing collagen-based hydrogels lack sufficient mechanical stability for controlled transfer post-thawing, making them unsuitable for regenerative medicine applications.
Innovation Solution
A method using a thin, biodegradable collagen cell carrier (CCC) with specific composition and mechanical properties, allowing cells to adhere and remain attached post-thawing, is developed. The CCC is air-dried to achieve high tensile strength and is used for seeding cells, which are then frozen and thawed without the need for protease treatment, maintaining biocompatibility and mechanical stability.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional cryopreservation methods are used with standard cell carriers, then cells can be frozen and stored, but cell vitality is significantly lost during freezing and thawing processes
Solution Approach 1:
The invention changes the physical parameters of the cell carrier by controlling its thickness to be between 10-150 μm and adjusting its water content to 40-80%. These parameter changes optimize heat transfer during freezing and thawing, reducing temperature gradients and minimizing cellular damage while maintaining structural integrity.
Solution Approach 2:
The invention uses collagen as the base material for the cell carrier, which provides both mechanical stability and biocompatibility. The collagen matrix forms a composite structure that protects cells during cryopreservation while maintaining their natural environment, thereby preserving cell vitality.
2Strength
If thicker cell carriers are used to provide mechanical stability, then structural support is improved, but temperature gradients increase during freezing and thawing which negatively affects cell survival
Solution Approach 1:
The invention optimizes the thickness parameter of the cell carrier to a specific range (10-150 μm) that balances mechanical stability with thermal conductivity. This thickness is sufficient to provide structural support for cell attachment while thin enough to allow rapid and uniform heat transfer during freezing and thawing, preventing damaging temperature gradients.
3Ease of operation
If conventional hydrogels are used as cell carriers, then cells can be cultured, but the carriers lack sufficient mechanical stability for controlled transfer after thawing
Solution Approach 1:
The invention uses collagen, a natural extracellular matrix protein, as the cell carrier material. Collagen provides inherent mechanical stability and structural integrity that allows the carrier to maintain its shape and provide controlled transfer capability after thawing, while still being biocompatible and supportive of cell culture.
4Quantity of substance
If cell carriers with high fluid absorption capacity are used, then cells can be supported, but crystallization occurs inside the carrier material during freezing
Solution Approach 1:
The invention controls the water content of the cell carrier to be within 40-80%, which is sufficient to support cell metabolism and culture requirements but limited enough to prevent excessive free water available for crystallization during freezing. This parameter control reduces internal ice crystal formation while maintaining 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 significantly increases cell survival rates and allows for the preservation of complex tissue structures, enabling their direct implantation in regenerative medicine by maintaining mechanical stability and biocompatibility throughout the cryopreservation process.
Implementation Method 1
The CCC is air-dried to achieve high tensile strength
Implementation Method 2
In order to protect cells during the freezing and thawing processes, special chemicals such as dimethyl sulfoxide (DMSO), trehalose, glycerine or hydroxyethyl starch (HES) are added to the freezing medium
Implementation Method 3
These so-called cryoprotectants protect the cells and their organelles from membrane damage by ice crystals and from intracellular dehydration during the transition of water from the liquid to the crystalline phase
Implementation Method 4
DMSO works primarily in the intracellular compartment by lowering the freezing point, which reduces the formation of ice crystals and thus minimizes cellular dehydration during freezing
Implementation Method 5
In contrast, the action of macromolecules such as hydroxyethyl starch is extracellular by forming a protective envelope for the cells and preventing loss of liquid from the cells during freezing
Data Source
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AI summary
The present invention belongs to the field of cryopreservation of cells and tissue cultures, more specifically it refers to a method for the cryopreservation and long term storage of cells, cell constructs or three-dimensional complex tissues assemblies. The methods are based on the use of a collagen cell carrier having a specific composition as well as to a very specific thickness and appropriate mechanical properties which are maintained after thawing. The use of such collagen cell carrier (CCC) provides a very suitable support for the cryopreservation resulting in very high survival rates after thawing and providing cells and tissue assemblies already adhered to a mechanically stable and biocompatible support. The invention also refers to the frozen collagen carrier-cells assembly and to the frozen artificial cell constructs or three-dimensional complex tissue assemblies obtainable by the method of the invention and to the use thereof after thawing.