Whole-Mount Cryopreservation for 3D Specimen Viability
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Solution Overview
Problem
Current cryopreservation technologies for 3D biological specimens, such as organoids and spheroids, suffer from high damage and loss during freezing and thawing processes due to complex handling steps, leading to low viability rates and inconsistent results, particularly in co-culture experiments.
Innovation Solution
A composition and associated device that allows whole-mount cryopreservation of biological specimens by maintaining them in their natural habitat, using cryoprotectant, diluting, and accessory agents, with controlled freezing and thawing, eliminating transfer and centrifugation steps.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional freezing and thawing processes are used for 3D biological specimens, then cryopreservation is achieved, but high damage and loss occur during transfer and handling steps
Solution Approach 1:
The patent combines the culture vessel and cryopreservation container into a single integrated system. The culture vessel serves dual purposes: as the growth environment during culture and as the cryopreservation container during freezing and storage. This eliminates the need to transfer specimens between different containers, thereby reducing handling steps and minimizing damage to 3D structures.
Solution Approach 2:
The culture vessel is designed to perform multiple functions: it serves as the incubation environment during cell growth, as the cryopreservation container during freezing, and as the storage vessel during long-term preservation. This multi-functionality eliminates the need for separate specialized containers for each stage, simplifying the overall process and reducing transfer-related damage.
2Reliability
If multiple transfer steps (from culture vessel to cryovial, centrifugation, pipetting) are performed, then cryopreservation is achieved, but specimen damage and data loss increase
Solution Approach 1:
The patent merges the culture vessel and cryopreservation container into one integrated system, eliminating the need to transfer specimens between different containers. This single-vessel system removes multiple transfer steps (from culture vessel to cryovial, centrifugation, pipetting) that cause specimen loss and damage, thereby improving preservation quality.
Solution Approach 2:
The patent extracts and eliminates unnecessary intermediate steps (centrifugation, pipetting, transfer to separate cryovials) from the cryopreservation process. By removing these harmful intermediate operations, the system directly reduces specimen loss while maintaining effective cryopreservation.
3Reliability
If 3D structures are transferred to separate cryovials, then cryopreservation is achieved, but 3D architecture and co-culture interactions are damaged
Solution Approach 1:
The patent combines the culture vessel and cryopreservation container into a single integrated system. The culture vessel serves dual purposes: as the growth environment during cell growth and as the cryopreservation container during freezing and storage. This eliminates the need to transfer specimens between different containers, thereby reducing handling steps and minimizing damage to 3D structures.
Solution Approach 2:
The patent performs preliminary action by preparing the cryopreservation medium and positioning all components within the culture vessel before initiating freezing. The vessel is pre-configured with appropriate sealing and cooling mechanisms, allowing the freezing process to begin directly without transfer, thus preserving 3D architecture and co-culture interactions.
4Reliability
If conventional cryopreservation methods are used, then cell viability is maintained, but time and labor for transfers and processing increase
Solution Approach 1:
The patent combines the culture vessel and cryopreservation container into a single integrated system. The culture vessel serves dual purposes: as the growth environment during cell growth and as the cryopreservation container during freezing and storage. This eliminates the need to transfer specimens between different containers, thereby reducing handling steps and minimizing damage to 3D structures.
Solution Approach 2:
The patent maintains continuity of useful action by keeping the culture vessel in place throughout the entire process: during active culture, during cryopreservation preparation, during freezing, and during long-term storage. This continuous presence eliminates interruption time from transfers and processing steps, reducing overall processing time 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 reduces specimen damage and loss, enhances viability, and ensures consistent preservation of 3D structures, facilitating accurate data retention and simplifying handling processes.
Implementation Method 1
the cells are treated with cryoprotectants and cooled down gradually to −196° C. Freezing cells slowly is essential to prevent intracellular ice formation
Implementation Method 2
cooled down gradually to −196° C.
Data Source
AI summary
The present invention relates to the compositions, methods, and associated devices (1) for the cryopreservation of biological specimens that enables to freeze and thaw the entire specimen by straightforwardly protecting its three-dimensional architecture. The invention comprises compositions including reagents to protect the specimen and preserve the physiological conditions. The methods comprise fewer steps excluding the need for other solutions used in conventional methods. The methods also eliminate stressful steps for biological specimens in conventional methods including harvesting, pipetting, centrifuging, and transferring. With this invention, the entire content of the specimen can be frozen, stored long-term in a single cell culture vessel with one of the associated devices (1) described here, and then thawed while still in the device (1). The biological specimens frozen by that method render high post-thawing viability. Moreover, the invention reduces cryoprotectant-related toxic events, human errors, and contamination risk.