Boron-Added Cryopreservation Medium for Stem Cell Viability
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Solution Overview
Problem
Current cryopreservation methods face challenges in maintaining cell viability and reducing stress during freezing, particularly due to the accumulation of ice crystals and toxic effects from cryoprotectants like dimethyl sulfoxide, which can lead to reduced cell survival and altered differentiation potential in stem cells.
Innovation Solution
A cryopreservation medium incorporating sodium pentaborate pentahydrate and reduced dimethyl sulfoxide concentrations is developed, which enhances cell viability and reduces stress during freezing, allowing for long-term storage of various cell types, including stem cells, while maintaining their multipotent properties.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If dimethyl sulfoxide (Me2SO) is used as a cryoprotectant during freezing, then cells are protected against ice crystal formation, but toxic effects accumulate reducing cell viability
Solution Approach 1:
The patent changes the chemical composition parameters of the cryopreservation medium by adding borate ions (sodium pentaborate pentahydrate) and adjusting Me2SO concentration. This parameter modification reduces the toxic effects of Me2SO while maintaining cryoprotective functionality, achieving cell viability above 80% after freeze-thaw cycles
Solution Approach 2:
Borate ions act as an intermediary substance that mitigates the harmful effects of Me2SO. The borate ions form complexes with Me2SO or modulate its activity, reducing toxicity while preserving the cryoprotective mechanism. This intermediary approach allows the system to maintain protection against ice crystals without the full burden of Me2SO toxicity
2Reliability
If freezing is performed slowly to prevent intracellular ice crystal formation, then cell structure is protected, but metabolites accumulate in the remaining liquid medium causing the solution effect
Solution Approach 1:
The patent modifies the chemical environment by introducing borate ions, which alter the freezing dynamics and metabolite behavior. The borate ions may chelate with metabolites or modify the eutectic point of the solution, reducing the concentration and harmful effects of accumulated metabolites during slow freezing
3Object-affected harmful factors
If Me2SO concentration is reduced to decrease toxicity, then cell viability increases, but cryoprotective effectiveness may be compromised
Solution Approach 1:
The patent creates a composite cryoprotective system combining Me2SO with borate ions (sodium pentaborate pentahydrate). This composite formulation allows for reduced Me2SO concentration while maintaining or enhancing cryoprotective effectiveness through the synergistic action of borate ions, which provide additional protection against ice crystal formation and membrane damage
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 medium effectively increases cell viability and reduces the toxic effects of dimethyl sulfoxide, enabling successful long-term storage and differentiation of stem cells, including mesenchymal and embryonic stem cells, with improved survival rates and preserved multipotency.
Implementation Method 1
There are cryoprotectants which are developed for this purpose and which protect the material that will be frozen against freeze-thaw stress
Implementation Method 2
The most important detrimental effect of the freezing process is accumulation of the water in the living system in intercellular and intracellular areas in the form of ice crystals due to decreasing of the temperature
Implementation Method 3
Chemicals such as glycerol, dimethyl sulfoxide (Me 2 SO) and 1,2 propandiol (PrOH) are used during freezing process and they replace water enabling it to flow out of the cell. This is used for eliminating the detrimental effects of the freezing process
Implementation Method 4
In order to stop all biological activity and realize the most effective biological freezing process, liquid nitrogen (-196°C) and vapor are preferred
Implementation Method 5
Another detrimental effect is the intracellular liquid flowing out of the cell and forming ice crystals in the intercellular area as a consequence of slow freezing application. Chemicals such as glycerol, dimethyl sulfoxide (Me 2 SO) and 1,2 propandiol (PrOH) are used during freezing process and they replace water enabling it to flow out of the cell. This is used for eliminating the detrimental effects of the freezing process (Stachecki et al., 1998 ). As much as the freezing process should be performed slowly, thawing process should be performed rapidly.
Data Source
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AI summary
The present invention relates to a cryopreservation (freezing) medium which can be used for long-term storage of cell lines, tissue samples, sperms, oocytes and embryos. The invention enables to freeze and store cells without affecting cell viability, to increase cell viability and to protect multipotent properties of the cells. The invention facilitates freezing and storing sperms, eggs, embryos, plant cells and materials, cancer and sensitive cell lines, stem cells (embryonic and mesenchymal), blood and blood cells and biological material and organs.