DMSO Cryopreservation Medium for Cell Survival During Freezing
Find Innovative SolutionsGenerate Solutions
Solution Overview
Problem
Cryopreservation of living cells is challenging due to harsh freezing and thawing conditions, leading to low survivability rates and cell damage, particularly from osmotic dehydration and intracellular freezing.
Innovation Solution
A cryopreservation medium comprising dimethyl sulfoxide (DMSO), disaccharides, human serum, and cytokines such as IL-7 or IL-15, with specific concentration ranges, is used to preserve mammalian cells, followed by controlled cooling and thawing.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Duration of action of stationary object
If cells are subjected to freezing and thawing conditions, then cells can be preserved for long-term storage, but cell survivability rate decreases due to harsh conditions
Solution Approach 1:
The patent uses a specialized cryopreservation medium containing DMSO, disaccharides, human serum, and cytokines as an intermediary substance between the cell and the freezing environment. This medium mediates the freezing process by controlling ice crystal formation, maintaining osmotic balance, and providing protective biochemical factors that reduce cellular stress during cryopreservation, thereby improving cell survivability while enabling long-term storage
Solution Approach 2:
The patent optimizes multiple parameters of the cryopreservation medium including DMSO concentration (1-10%), disaccharide concentration (0.25-5%), human serum concentration (10-90%), and cytokine concentrations (IL-7 at 1-50 ng/mL, IL-15 at 1-50 ng/mL). By systematically adjusting these chemical and physical parameters, the medium creates optimal conditions that protect cells during freezing and thawing while maintaining long-term storage capability
2Temperature
If extracellular medium freezes, then cells can be preserved at low temperature, but intracellular water loss increases leading to osmotic dehydration and cell damage
Solution Approach 1:
Disaccharides in the cryopreservation medium act as protective intermediaries that bind to cell membranes and proteins, stabilizing their structure during freezing. They also regulate osmotic pressure by forming a protective matrix that prevents excessive water efflux from cells, thereby reducing osmotic dehydration damage while enabling freezing preservation
Solution Approach 2:
The cytokines IL-7 and IL-15 are added to the cryopreservation medium beforehand to activate cellular protective pathways and enhance cell resistance to freezing stress. This pre-conditioning provides a cushioning effect that protects cells from osmotic dehydration damage before the actual freezing occurs, improving survivability without compromising the freezing temperature requirement
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 results in high survivability of thawed cells, suitable for various applications including adoptive cell transfer, with enhanced cell survival both in vitro and post-transplantation.
Implementation Method 1
dimethyl sulfoxide (DMSO)...allows for freezing and subsequent thawing of viable cells
Implementation Method 2
cells attempt to maintain osmotic equilibrium across the membrane leading to intracellular water loss
Implementation Method 3
disaccharide, human serum...retain high survivability
Data Source
AI summary
The present disclosure provides, among other things, a cryopreservation medium for cryopreserving mammalian cells, the medium comprising: dimethyl sulfoxide (DMSO), disaccharide, human serum, and IL-7 and/or IL-15. The present disclosure also provides, among other things, a cryopreservation medium for cryopreserving mammalian cells, the medium comprising: between about 1 w/v % and 10 w/v % dimethyl sulfoxide (DMSO), between about 0.25 w/v % and 5 w/v % disaccharide, and between about 10 w/v % and 90 w/v % human serum. The present disclosure also provides, among other things, a cryopreservation medium for cryopreserving mammalian cells, the medium comprising: between about 1 w/v % and 10 w/v % dimethyl sulfoxide (DMSO), between about 0.25 w/v % and 5 w/v % disaccharide, and between about 0.5 w/v % and 30 w/v % human serum albumin.