Cryopreservation Using 3-OMG Ice Nucleation
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Current methods for preserving biological samples, organs, and organisms face challenges such as limited success in preserving complex systems, with hypothermic storage being inadequate and vitrification involving toxic cryoprotectants and osmotic damage, while supercooling methods are thermodynamically unstable and prone to freezing in larger tissues.
Innovation Solution
The method involves using 3-O-methyl-D-glucopyranose (3-OMG) as an ice nucleating agent to uniformly nucleate ice across the microvasculature, reducing intracellular ice formation, and employing a combination of cryoprotectants like polyethylene glycol (PEG) and glycerol in a controlled freezing protocol to achieve partial freezing, allowing for longer preservation times without extensive tissue damage.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Duration of action of stationary object
If vitrification is used to preserve complex systems, then preservation duration is extended, but toxicity from high molarity cryoprotectants and osmotic damage occur
Solution Approach 1:
The patent changes the fundamental parameter of ice formation from avoided (vitrification) to controlled and uniform (heterogeneous nucleation). By introducing ice nucleating agents that promote uniform extracellular ice formation, the method achieves long-term preservation without requiring high molarity cryoprotectants, thus avoiding their toxic effects and osmotic damage.
Solution Approach 2:
The patent introduces ice nucleating agents as intermediaries that facilitate controlled ice formation. These agents act as mediators between the preservation system and ice crystallization, enabling uniform extracellular ice formation that protects cells from intracellular ice damage without requiring toxic high concentrations of cryoprotectants.
2Duration of action of stationary object
If supercooling is used to extend preservation duration, then preservation time is tripled, but thermodynamic instability and freezing probability increase
Solution Approach 1:
The patent applies preliminary action by introducing ice nucleating agents before cooling begins. These agents pre-establish nucleation sites that trigger controlled ice formation at higher temperatures, preventing supercooling and its associated thermodynamic instability. This preliminary nucleation ensures reliable preservation without the freezing risks of supercooling.
3Reliability
If uniform ice nucleation is achieved using ice nucleating agents, then intracellular ice formation is reduced, but preservation protocol complexity increases
Solution Approach 1:
The patent employs inexpensive ice nucleating agents (such as bacteria or proteins) that can be easily added to the preservation solution. These simple, low-cost additives provide the necessary nucleation function without requiring complex equipment or elaborate protocols, making the method both reliable and practical for clinical 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 approach significantly improves endothelial cell viability and attachment during preservation, maintaining cell membrane integrity and function, and can extend preservation periods beyond traditional methods, achieving a thermodynamically stable frozen state at colder temperatures without the drawbacks of vitrification.
Implementation Method 1
incubating the cell in a loading solution comprising 3-O-methyl-D-glucopyranose (3-OMG) in a vessel; cooling the cell in the vessel to a temperature from −5° C. to −40° C., thereby partially freezing the vessel
Implementation Method 2
cooling the cell in the vessel to a temperature from −5° C. to −40° C., thereby partially freezing the vessel, wherein from about 5% to about 95% of the storage solution in the vessel is in solid state
Data Source
AI summary
This disclosure is related to methods of preserving biological samples, organs. and organisms.