Cryopreservation Cell Holder with Filter for Vitrification
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Solution Overview
Problem
Current cryopreservation methods for biological cells and tissues, such as oocytes and embryos during IVF procedures, are time-consuming, require high skill, and involve significant cell manipulation, with permeating cryoprotectants posing toxicity risks due to their high concentrations needed for rapid cooling and vitrification.
Innovation Solution
A method and apparatus for cryopreservation using a cell holder with a filter and multiple treatment stations for controlled washing and rapid cooling, allowing for simultaneous processing of multiple cells, minimizing manipulation, and employing a sequence of treatment solutions to achieve vitrification without prolonged exposure to toxic cryoprotectant concentrations.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If permeating cryoprotectants are used at high concentrations for rapid cooling and vitrification, then ice crystal formation is prevented and vitrification is achieved, but cell toxicity increases significantly
Solution Approach 1:
The patent divides the cryoprotectant application into distinct stages: a first solution with lower concentration applied first, followed by a second solution with higher concentration. This segmentation allows the cell to gradually adapt to increasing cryoprotectant concentrations, reducing toxic shock while ultimately achieving the high concentration needed for successful vitrification and ice crystal prevention.
Solution Approach 2:
The patent applies a preliminary lower concentration of permeating cryoprotectant before applying the higher concentration. This preliminary action prepares the cell membrane and internal environment for the subsequent high-concentration exposure, reducing the harmful toxic effects while ensuring the cell is ready for effective vitrification when the higher concentration is applied.
2Reliability
If traditional slow cooling method is used to initiate ice crystal formation remotely, then ice crystals form away from cells, but the process is time-consuming and requires precise temperature control
Solution Approach 1:
The patent utilizes the phase transition of water to glass (vitrification) by applying extremely rapid cooling rates. Instead of allowing controlled ice crystal formation through slow cooling, the method transitions the aqueous solution directly into a vitrified glass-like state, preventing ice crystal formation entirely while dramatically reducing the cooling time required for cryopreservation.
3Reliability
If manual manipulation and washing of cells through multiple solutions is performed, then cells are properly prepared for cryopreservation, but the process requires high skill and is time-consuming
Solution Approach 1:
The patent introduces a nonpermeating cryoprotectant as an intermediary substance that facilitates the washing and preparation process. This intermediary agent helps control osmotic pressure and cell dehydration during the transition between different solutions, making the manual manipulation process more forgiving and less skill-dependent while maintaining high preparation quality.
Solution Approach 2:
The patent systematically changes solution parameters including composition, concentration, and temperature across multiple washing steps. By carefully controlling these parameter transitions, the method standardizes the cell preparation process, reducing variability and making it more reproducible with less operator skill while maintaining high reliability.
4Reliability
If cells are exposed to high concentration permeating cryoprotectant for prolonged periods, then adequate protection against ice crystals is achieved, but metabolic damage occurs due to toxicity
Solution Approach 1:
The patent segments the exposure to high-concentration permeating cryoprotectant into brief, controlled intervals separated by exposure to lower concentration solutions. This segmented approach allows the cell to receive sufficient protective effect from the high concentration while limiting the cumulative toxic exposure time, preventing metabolic damage while maintaining ice crystal protection.
Solution Approach 2:
The patent employs periodic action by alternating between solutions of different cryoprotectant concentrations during the preparation process. Cells are exposed to high concentration for brief protective intervals, then rested in lower concentration solutions, creating a periodic pattern that delivers necessary protection while minimizing toxic exposure duration and metabolic 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 method enables efficient, safe, and hygienic cryopreservation with reduced toxicity exposure, allowing for high survival rates of cells by maintaining precise control over the washing and cooling processes, and facilitating automation for increased efficiency.
Implementation Method 1
a filter to retain the organic cell whilst allowing passage of treatment solution
Implementation Method 2
a more recent method known as vitrification, transforms the solution into a glass-like amorphous solid that is free from any crystalline structure, following extremely rapid cooling
Implementation Method 3
The cell and treatment solution are rapidly cooled to a predetermined cryopreservation temperature sufficient to solidify the treatment solution
Implementation Method 4
They form hydrogen bonds with water molecules and prevent ice crystallization
Implementation Method 5
prevent ice crystallization
Implementation Method 6
They act by drawing free water from within the cell, thus dehydrating the intracellular space
Implementation Method 7
at high enough concentrations, they inhibit the formation of the characteristic ice crystal and lead to the development of a solid, glasslike, or vitrified state in which water is solidified, but not crystalline or expanded
Data Source
Figure 1a~1f
Figure 2a~2d
Figure 3a~3c
AI summary
The method involves placing an oocyte cell in a cell holder (1), securing the cell holder in a treatment station, applying a treatment solution to said cell by washing the cell with the solution, and rapidly cooling the cell holder and cell to a predetermined cryopreservation temperature for cryopreservation of the cell. The cell is cooled at a high rate sufficient to permit vitrification of the cell and any surrounding treatment solution to occur. The cell is then maintained at or below a predetermined storage temperature for storage. The method allows multiple cells to be treated simultaneously each secured within a respective cell holder.