Zwitterionic Cryoprotectant Composition for Toxicity-Free Cell Preservation
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Solution Overview
Problem
Current cell cryopreservation technologies face challenges due to the toxicity of conventional cryoprotectants like DMSO and glycerol, which can cause cell damage and side effects, and require cumbersome stepwise cooling procedures, leading to inefficient and costly cryopreservation processes.
Innovation Solution
A zwitterionic molecule-based cell cryopreservation composition, excluding toxic substances like DMSO and glycerol, is used for ultrarapid cryopreservation, providing a non-toxic and efficient method that allows for direct cell use after recovery without extensive washing, utilizing a nutrient-rich solution with zwitterionic molecules to regulate osmotic pressure and lower the freezing point.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If conventional cryoprotectants like DMSO and glycerol are used, then cell cryopreservation can be achieved, but cell toxicity and side effects occur
Solution Approach 1:
The patent changes the chemical composition parameters by replacing toxic conventional cryoprotectants (DMSO, glycerol) with non-toxic zwitterionic compounds. This substitution maintains the cryoprotective function while eliminating cellular toxicity, directly resolving the contradiction between preservation effectiveness and cell safety
Solution Approach 2:
The patent employs a simplified single-step ultra-rapid freezing protocol that eliminates the need for prolonged exposure to toxic cryoprotectants. By using non-toxic zwitterionic compounds that allow immediate freezing without extended contact time, the method dispenses with the need for extensive washing steps and multiple handling procedures, thereby eliminating toxicity while maintaining preservation efficacy
2Reliability
If stepwise cooling procedures are used with conventional cryoprotectants, then cell preservation can be achieved, but the process becomes cumbersome and inefficient
Solution Approach 1:
The patent implements ultra-rapid freezing that skips the traditional multi-stage cooling process. By using non-toxic zwitterionic compounds, the method enables direct ultra-rapid freezing without requiring gradual temperature reduction steps, dramatically shortening the cryopreservation time from hours to minutes while maintaining high cell survival rates
Solution Approach 2:
The patent applies zwitterionic compounds to cells before freezing to prepare them for ultra-rapid temperature changes. This preliminary treatment with non-toxic compounds protects cells during the rapid freezing process, eliminating the need for subsequent washing steps and multiple handling operations, thereby improving operational efficiency
3Reliability
If conventional cryoprotectants are used, then cell freezing can be achieved, but multiple refrigeration devices and extensive washing are required
Solution Approach 1:
The patent extracts and eliminates the need for multiple refrigeration devices by using non-toxic zwitterionic compounds that enable ultra-rapid freezing in a single step. The method removes the requirement for sequential use of different temperature chambers (−80°C, −20°C, 4°C), simplifying the equipment infrastructure while maintaining preservation effectiveness
Solution Approach 2:
The patent employs a universal non-toxic zwitterionic compound formulation that works across different cell types and concentrations. This single composition handles both small and large volume samples without requiring different protocols or equipment, eliminating the need for multiple specialized refrigeration devices and simplifying the overall cryopreservation system
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 zwitterionic molecule composition achieves high cell survival rates and simplifies the cryopreservation process, reducing the need for multiple refrigeration devices and minimizing cell damage, making it suitable for various cell types and concentrations.
Implementation Method 1
utilizing a nutrient-rich solution with zwitterionic molecules to regulate osmotic pressure and lower the freezing point
Implementation Method 2
utilizing a nutrient-rich solution with zwitterionic molecules to regulate osmotic pressure and lower the freezing point
Data Source
AI summary
The present application relates to a cell cryopreservation protective composition, the use of the composition, and a cell cryopreservation method. The cell cryopreservation protective composition comprises a zwitterionic molecule having the general formula R1—N+(CH3)2—(CH2)n—R2, wherein R1 is a linear or branched alkyl having 1 to 10 carbon atoms, and is optionally substituted with a substituent; R2 is any negatively charged group selected from the group consisting of —COO−, —SO4−, —SO3−, and (I); and R3 is a group selected from the group consisting of (methyl)acryloyloxyalkyl, alkyl and alkenyl; and the zwitterionic molecule having general formula R1—N+(CH3)2—(CH2)n—R2 is preferably a betaine compound. The cell cryopreservation protective composition can carry out cell cryopreservation in a non-toxic and efficient manner, and results in an extremely high post-thaw cell survival rate and does not require stepwise cryopreservation. After cell recovery, the cells can be used directly or after being slightly diluted.