Cryopreservation Liquid Composition for Biological Materials
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Solution Overview
Problem
Current vitrification protocols for cryopreserving biological materials are complex, lack standardization, and often use undefined components, leading to instability, safety concerns, and reduced reproducibility.
Innovation Solution
A chemically defined liquid composition comprising a polar aprotic solvent, a monohydric or polyhydric alcohol, an unbranched polysaccharide, a branched polysaccharide, and polyvinyl alcohol, which provides a synergistic effect for optimal cryopreservation characteristics.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If current vitrification protocols are used, then cryopreservation efficiency is improved, but methodological complexity and lack of standardization increase
Solution Approach 1:
The patent applies parameter changes by precisely defining the concentrations and compositions of cryoprotectants in the vitrification solution. The solution contains DMSO (10-20% v/v), ethylene glycol (5-15% v/v), sucrose (5-20% w/v), and dextran (0.1-1% w/v), with each component's concentration optimized to achieve reliable cryopreservation while simplifying the protocol. This standardized compositional approach resolves the contradiction by maintaining high efficiency through controlled parameters while reducing methodological complexity.
Solution Approach 2:
The patent employs composite materials by combining multiple cryoprotectants (DMSO, ethylene glycol, sucrose, dextran) in a specifically formulated vitrification solution. This composite approach leverages the synergistic effects of different substances: DMSO and ethylene glycol as permeating cryoprotectants, sucrose and dextran as non-permeating agents. The composite formulation achieves reliable cryopreservation outcomes while providing a standardized, reproducible protocol that reduces methodological complexity.
2Reliability
If undefined components from human or animal fluids are used, then biological activity is improved, but biological safety and reproducibility deteriorate
Solution Approach 1:
The patent applies this principle by replacing expensive, undefined biological components (such as fetal bovine serum or other animal-derived additives) with well-defined, chemically characterized substances. The vitrification solution uses DMSO, ethylene glycol, sucrose, and dextran—all of which have known purities, stable properties, and established safety profiles. This substitution maintains necessary biological activity while eliminating the biological safety risks and reproducibility issues associated with undefined components from human or animal fluids.
Solution Approach 2:
The patent applies local quality by ensuring that each component in the vitrification solution has a clearly defined function and property profile. DMSO and ethylene glycol are specified for their permeating cryoprotective properties, sucrose for its non-permeating osmotic protection, and dextran for its viscosifying and protective effects. This precise specification of local qualities (functions and properties of individual components) ensures biological activity while guaranteeing biological safety and reproducibility through well-characterized materials.
3Reliability
If multiple cryoprotectants are combined, then cryopreservation efficacy is improved, but intracellular ingress of cryoprotectants increases
Solution Approach 1:
The patent applies parameter changes by carefully controlling the concentrations of permeating cryoprotectants (DMSO at 10-20% v/v and ethylene glycol at 5-15% v/v) in the vitrification solution. These concentrations are optimized to achieve sufficient intracellular protection without excessive ingress that could cause toxicity. The balanced formulation, combined with controlled cooling rates and exposure times, maintains high cryopreservation efficacy while minimizing harmful intracellular accumulation of cryoprotectants.
Solution Approach 2:
The patent employs composite materials by combining permeating cryoprotectants (DMSO, ethylene glycol) with non-permeating agents (sucrose, dextran) in a vitrification solution. This composite approach allows the permeating agents to provide intracellular protection at lower, safer concentrations, while the non-permeating agents contribute to extracellular osmotic balance and vitrification. The synergistic interaction in this composite system achieves high cryopreservation efficacy while reducing the total quantity of permeating cryoprotectants that would otherwise need to be used at higher concentrations.
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 ensures high survival and hatching rates of cryopreserved biological materials, including stem cells, embryos, and gametes, while maintaining biological safety and reproducibility across various cryopreservation procedures.
Implementation Method 1
The VIT procedures were introduced as an alternative to SLF in order to reduce the likelihood of intracellular ice crystal formation
Implementation Method 2
Cryopreservation is amongst the most powerful and efficient tools for indefinitely preserving the genetics of economically, therapeutically or scientifically important cells, embryos or gametes
Implementation Method 3
Since water is the major component of the cell, its solidification must be tightly controlled during both cooling and subsequent warming to avoid intracellular formation of ice crystals
Data Source
AI summary
The present application discloses a liquid composition for cryopreservation of a biological material comprising a polar aprotic solvent, a monohydric or polyhydric alcohol, an unbranched polysaccharide, a branched polysaccharide; and polyvinyl alcohol. Further provided are uses of said liquid composition for cryopreservation of abiological material and methods for preserving a biological material using said liquid composition.