Mitochondria Cryopreservation Composition for Viability and Long-Term Storage
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Solution Overview
Problem
Existing cryopreservation methods for mitochondria result in damage and dysfunction due to ice crystal formation, osmotic shock, and membrane damage during freezing and thawing, limiting their storage time and viability for research and therapeutic applications.
Innovation Solution
A composition comprising an aqueous buffer with a pH of 5.5 to 8.5, trehalose at a concentration of at least 150 mM, and a combination of cryoprotecting agents such as amino acids, sugars, and polymers, including specific concentrations of trehalose, amino acids like proline, and polymers like polyethylene glycol, to maintain mitochondrial integrity and function during cryopreservation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Duration of action of stationary object
If conventional cryopreservation methods are used for mitochondria, then storage is possible, but mitochondrial damage occurs due to ice crystal formation, osmotic shock, and membrane damage
Solution Approach 1:
The invention changes the chemical composition parameters of the cryopreservation medium by incorporating specific concentrations of trehalose (at least 150 mM), amino acids (at least 150 mM total concentration), and optional polymers (2.5% to 30% w/v). These parameter modifications create an optimized chemical environment that prevents ice crystal formation and osmotic shock, thereby maintaining mitochondrial viability during long-term storage.
Solution Approach 2:
The invention uses a composite cryoprotective system combining multiple substances: trehalose as a disaccharide, amino acids (such as proline), and optional polymer additives. This composite approach provides synergistic protection where each component addresses different aspects of cryodamage, collectively maintaining mitochondrial structure and function during freezing and thawing.
2Reliability
If cryoprotective agents are added to preserve mitochondria during freezing, then mitochondrial integrity is maintained, but the composition complexity increases
Solution Approach 1:
The invention establishes specific concentration parameters for each cryoprotective component (trehalose at least 150 mM, amino acids at least 150 mM total) to achieve optimal protection. By defining precise parameter ranges, the invention simplifies the formulation process while ensuring reliable mitochondrial preservation, reducing the need for complex trial-and-error optimization.
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
The composition effectively preserves mitochondrial structure and function, enabling long-term storage and ready-to-use mitochondria for therapeutic applications, such as mitochondrial transplantation, by minimizing damage and maintaining bioenergetic functions.
Implementation Method 1
The present application relates to compositions and methods for the cryopreservation of compositions comprising isolated viable mitochondria
Implementation Method 2
Existing cryopreservation methods for mitochondria result in damage and dysfunction due to ice crystal formation, osmotic shock, and membrane damage during freezing and thawing
Implementation Method 3
an aqueous buffer having a pH of 5.5 to 8.5
Data Source
AI summary
The disclosure relates to composition and method for cryopreservation of mitochondria, cryopreserved composition comprising mitochondria and their therapeutic use.