Non-linear Cooling Cryopreservation Protocol for Stem Cells
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Solution Overview
Problem
Current cryopreservation methods for cells and tissues face challenges in achieving optimal recovery rates due to injury during the freezing process, particularly with the use of permeating cryoprotectants like DMSO, and lack a reliable method for optimizing cooling profiles without toxic cryoprotectants.
Innovation Solution
A non-linear cooling cryopreservation method is developed, which determines an optimal cooling profile using computer simulations based on cellular osmotic transport properties and thermodynamic parameters to minimize intracellular supercooling and ice nucleation, allowing for maximum cell recovery without the need for toxic cryoprotectants.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If permeating cryoprotectants like DMSO are used for cryopreservation, then cell recovery is improved, but adverse effects and toxicity increase
Solution Approach 1:
The patent extracts and removes the harmful permeating cryoprotectant (DMSO) from the cryopreservation system while maintaining cell recovery through alternative means - specifically through optimized non-linear cooling profiles that control ice crystal formation and osmotic stress without requiring toxic chemical additives
Solution Approach 2:
The patent changes the physical parameters of the freezing process by implementing non-linear cooling profiles with specific cooling rates that vary over time, transforming the cryopreservation approach from chemical-based protection to physically-controlled temperature management that avoids toxicity while maintaining effectiveness
2Ease of operation
If constant cooling rate is used for cryopreservation, then process simplicity is maintained, but cell recovery is suboptimal due to freezing injury
Solution Approach 1:
The patent applies dynamics by transitioning from a static constant cooling rate to a dynamic non-linear cooling profile where the cooling rate changes over time according to specific mathematical functions, allowing the system to adapt to different stages of freezing and minimize cell injury while maintaining operational feasibility
Solution Approach 2:
The patent implements preliminary action by pre-calculating and pre-programming the optimal non-linear cooling profile based on cell type and cryoprotectant characteristics, so that the optimized cooling schedule is established before the actual freezing process begins, eliminating the need for complex real-time adjustments during freezing
3Reliability
If cooling rate is optimized for specific cell types, then cell recovery is improved, but process complexity and optimization time increase
Solution Approach 1:
The patent changes key parameters of the cooling profile (cooling rate, hold temperature, hold time) based on cell-specific characteristics, allowing optimization for different cell types while maintaining a systematic approach that reduces overall complexity through standardized parameter adjustment rather than complete re-optimization
Solution Approach 2:
The patent uses copying by creating standardized cooling profile templates for different cell types based on their general characteristics, allowing rapid deployment of optimized protocols without requiring extensive experimentation for each new cell application, thus reducing optimization complexity while maintaining effectiveness
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
This method achieves comparable recovery rates to those using cryoprotectants, with up to 75% recovery of cells preserved without cryoprotectants, and can be applied to various cell types, including stem cells, by controlling temperature and hold times to prevent intracellular freezing and osmotic stress.
Implementation Method 1
cooling the cells to a first temperature for a first period of time, then cooling the cells to a storage temperature
Implementation Method 2
determining an optimal cooling profile for maximum recovery of the cells... based on cellular osmotic transport properties
Data Source
AI summary
A non-linear cooling cryopreservation method for improving cryopreservation protocols for cells that involves producing a simulation of cellular responses to a range of cooling parameters; determining optimal cooling parameters required to minimize cryoinjury to the cells using simulation of cellular responses and experimental results; and incorporating optimal parameters into the protocol. The simulation is based on mathematical models of cellular parameters. A non-linear cooling cryopreservation protocol for cryopreserving stem cells is also disclosed that does not require cryoprotectants.