Mitochondria Extraction Kit with Composite Storage Solution
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Solution Overview
Problem
Current methods for extracting and storing mitochondria for transplantation are limited by the difficulty in maintaining mitochondrial function and purity, with existing kits lacking transparency in their formulations and no effective domestic alternatives available.
Innovation Solution
A kit and method for extracting cell mitochondria in vitro, comprising specific solutions (extracting solutions A, B, and C, and a storage solution) that include EDTA, sodium lactate, mannitol, heparin, and other components, designed to maintain mitochondrial function and viability.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Ease of manufacture
If chemical lysis method is used for mitochondrial extraction, then the process does not require additional instruments and concentration is controllable, but the composition of extraction reagents is not disclosed and there are no effective domestic alternatives
Solution Approach 1:
The extraction process is divided into multiple stages using different solutions: extracting solution A (for cell lysis), extracting solution B (for mitochondrial isolation), and extracting solution C (for purification). Each solution has a specific function and composition, allowing systematic optimization of each step while maintaining overall process simplicity.
Solution Approach 2:
The patent specifies precise concentrations and compositions for each extraction solution (e.g., EDTA 1-10 mM, sodium lactate 10-50 mM, mannitol 200-800 mM), transforming the vague chemical lysis approach into a controlled multi-parameter process that optimizes both ease of operation and reagent transparency.
2Duration of action of stationary object
If mitochondria are stored in conventional formulation, then they can be stored, but they lose membrane potential and respiratory function in a relatively short period of time
Solution Approach 1:
The storage solution is formulated as a composite mixture containing multiple components working together: mannitol (200-800 mM) for osmotic protection, EDTA (1-10 mM) for metal chelation, sodium lactate (10-50 mM) for metabolic support, succinic acid (10-50 mM) and sodium pyruvate (10-50 mM) for respiratory chain maintenance, and Tris-HCl or HEPES (10-50 mM) for pH buffering. This composite formulation synergistically maintains membrane potential and respiratory function during storage.
Solution Approach 2:
The storage solution optimizes multiple parameters simultaneously: osmotic pressure (via mannitol), pH (via Tris-HCl or HEPES), metal ion chelation (via EDTA), and metabolic substrates (via succinic acid and sodium pyruvate). This multi-parameter optimization extends storage duration while maintaining mitochondrial function.
3Reliability
If grinding homogenization is used for cell lysis, then it is the earliest method applied, but it requires high precision and specific grinding instruments which is not conducive to direct clinical use
Solution Approach 1:
The patent replaces mechanical grinding homogenization with chemical lysis using a defined combination of solutions (extracting solution A, B, and C). This substitution eliminates the need for complex mechanical instruments while maintaining effective cell lysis and mitochondrial isolation through controlled chemical actions.
4Ease of manufacture
If ultrasonic lysis is used for cell lysis, then it can lyse cells, but it generates additional heat and the long-term process will affect the viability of mitochondria
Solution Approach 1:
The patent replaces ultrasonic mechanical lysis with chemical lysis using extracting solution A (containing EDTA and sodium lactate). This substitution avoids heat generation entirely while maintaining effective cell lysis, thereby preserving mitochondrial viability.
Solution Approach 2:
The patent converts the potential harm of heat generation into a benefit by using chemical lysis that operates at ambient temperatures. The chemical solutions (particularly EDTA and sodium lactate) provide effective lysis without thermal effects, turning the temperature control issue into an advantage for maintaining mitochondrial function.
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 proposed solution effectively maintains mitochondrial function and viability, prolonging the interval time from extraction to application, and facilitating the clinical application of mitochondrial transplantation.
Implementation Method 1
calcium ion chelators (such as EGTA, EDTA)
Implementation Method 2
chemical lysis method, including the use of digitonin or heparin, destroys the structure of the outer cell membrane and causes it to release cellular components
Implementation Method 3
osmotic pressure regulating solutions (such as mannitol, sorbitol, sucrose)
Implementation Method 4
mitochondria preserved with the current formulation will lose membrane potential and respiratory function
Implementation Method 5
ATP-producing capacity
Implementation Method 6
ion buffers (such as Tris-Hcl, HEPES)
Data Source
AI summary
The invention provides a kit and a method for extracting cell mitochondria in vitro. The kit comprises an extracting solution A, an extracting solution B, an extracting solution C and a storage solution, wherein the extracting solution A comprises EDTA (Ethylene Diamine Tetraacetic Acid), sodium lactate and sodium chloride, the extracting solution B is a heparin solution, the extracting solution C comprises mannitol, EDTA, sodium lactate and sodium chloride, and the storage solution comprises mannitol, EDTA, sodium lactate, succinic acid, sodium pyruvate and sodium chloride. The functions of mitochondria can be better maintained in vitro by utilizing the extracting solution and the storage solution, the interval time from mitochondria extraction to mitochondria application to the body of a patient is prolonged, and beneficial help is provided for clinical application of mitochondria transplantation.