Porous Membrane Cryopreservation for Long-Term Cell Storage
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Solution Overview
Problem
Existing cryopreservation methods using liquid nitrogen are costly, complex, and limited to short-term preservation, while methods at higher temperatures stress cells and require complicated operations, and there is a lack of effective long-term preservation techniques focusing on the cell culture substrate.
Innovation Solution
Cryopreservation of cells on a porous membrane, particularly a polymer porous membrane, at temperatures above -150°C without liquid nitrogen, allowing for long-term preservation and simplified operations.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Duration of action of stationary object
If liquid nitrogen is used for cryopreservation, then cells can be preserved for long periods, but the cost increases and management becomes difficult
Solution Approach 1:
The patent changes the temperature parameter from cryogenic (-196°C with liquid nitrogen) to higher temperatures (-80°C to -20°C) achievable with ordinary freezers. This parameter change eliminates the need for liquid nitrogen while maintaining cell viability, resolving the contradiction between preservation duration and management complexity
Solution Approach 2:
The patent replaces expensive liquid nitrogen with ordinary freezing agents (ice, salt, or simple refrigeration) that are inexpensive and easily managed. This substitution maintains adequate preservation capability while eliminating the complexity of liquid nitrogen management
2Duration of action of stationary object
If cells are cryopreserved in suspension state, then preservation is possible, but complicated operations including washing and centrifugation are required
Solution Approach 1:
The patent performs preliminary attachment of cells to a substrate (such as a culture dish or membrane) before cryopreservation. This preliminary action eliminates the need for subsequent washing and centrifugation steps during preservation, as cells remain fixed in place throughout the freezing process
Solution Approach 2:
The patent combines the cell attachment function with the preservation container by using culture dishes or membranes as the preservation substrate. This merging eliminates the need for separate handling steps and simplifies the overall preservation process
3Temperature
If cells are cryopreserved at -80°C, then short-term preservation is possible, but long-term preservation cannot be achieved
Solution Approach 1:
The patent optimizes the temperature parameter within the -80°C to -20°C range and combines it with optimized preservation solutions and cell attachment methods. This parameter optimization enables long-term preservation without requiring liquid nitrogen temperatures, resolving the contradiction between temperature and preservation duration
4Ease of operation
If trypsin treatment and washing are performed for cell preservation, then cells can be collected, but stress is applied to cells
Solution Approach 1:
The patent performs preliminary attachment of cells to a substrate before preservation, eliminating the need for trypsin treatment and washing. Cells are directly frozen in their attached state, avoiding mechanical and chemical stress while maintaining ease of operation
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
Enables stable long-term preservation of cells without impairing their function, reducing costs by eliminating the need for liquid nitrogen and simplifying operations, facilitating long-distance transportation and easy cell culture post-thawing.
Implementation Method 1
Cryopreservation of cells on a porous membrane, particularly a polymer porous membrane, at temperatures above -150°C without liquid nitrogen
Data Source
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AI summary
The present invention provides a method for cryopreserving cells, including cryopreserving cells for 30 days or more in a state in which the cells are seeded on a porous membrane without using liquid nitrogen. The present invention further provides a cell culture method including a step of cryopreserving cells seeded on a porous membrane for 30 days or more without using liquid nitrogen. Further, the present invention provides a cryopreservation substrate formed of a porous membrane for use in cryopreserving seeded cells for 30 days or more.