Viable Cell Cryopreservation Medium with DMSO and HES
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Solution Overview
Problem
Current methods for cryopreserving viable cells face challenges in maintaining viability and stability during storage and distribution, requiring effective additives that are biologically acceptable and cost-effective for broad application in therapies.
Innovation Solution
A method involving a cryopreservation medium with dimethylsulfoxide at 2% and hydroxyethyl starch at 6% by weight, combined with water, which is cooled to -60°C to -100°C for cryopreservation and then diluted with an aqueous medium to reduce DMSO concentration to 0.5% or less, maintaining cell viability and stability for extended storage and administration.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If high concentrations of DMSO are used for cryopreservation, then cell viability during storage is improved, but biological acceptability and safety for patient administration deteriorates
Solution Approach 1:
The patent changes the concentration parameter of DMSO from traditional high levels (10-20%) to a low level (2%), and compensates by adjusting the concentration of HES from 1-5% to 6%. This parameter substitution resolves the contradiction by maintaining cryoprotection effectiveness while eliminating the harmful effects of high DMSO concentrations.
Solution Approach 2:
The patent creates a composite cryopreservation system combining DMSO and HES in specific proportions (2% DMSO + 6% HES). This composite approach allows the weaker DMSO concentration to work synergistically with HES to provide adequate cryoprotection without the harmful effects of high DMSO alone.
2Reliability
If expensive cryogenic storage infrastructure is used, then cell storage stability is improved, but manufacturing and distribution cost increases
Solution Approach 1:
The patent changes the storage temperature parameter from ultra-low cryogenic temperatures (-196°C liquid nitrogen) to moderately low temperatures (-60°C to -100°C mechanical freezer). This parameter change enables the use of standard mechanical freezers instead of expensive cryogenic infrastructure, reducing costs while maintaining storage stability.
Solution Approach 2:
The patent employs a cryopreservation formulation (2% DMSO + 6% HES) that enables storage in disposable or non-reusable mechanical freezers rather than requiring permanent, expensive cryogenic infrastructure. This makes the system more accessible and cost-effective for broad distribution.
3Object-affected harmful factors
If DMSO concentration is reduced for safety, then biological acceptability is improved, but cell viability during cryopreservation deteriorates
Solution Approach 1:
The patent changes the concentration parameter of HES from 1-5% to 6%, which is slightly higher than the lower end of the traditional range. This increased HES concentration compensates for the reduced DMSO concentration, maintaining cryoprotection effectiveness while improving biological acceptability.
Solution Approach 2:
The patent introduces HES as an intermediary substance that mediates between the need for cryoprotection and biological safety. HES acts as a substitute that can partially replace DMSO's protective function while being more biologically acceptable, allowing low DMSO concentrations to be used safely.
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 approach allows for robust, storable, and viable cell preparations that retain high cell viability during storage and upon thawing, enabling efficient distribution and therapeutic use without the need for expensive cryogenic storage, with the cells maintaining at least 80% viability after six months at -60°C to -100°C.
Implementation Method 1
thawing a cryopreserved composition that includes viable cells and an aqueous cryopreservation medium, wherein the aqueous cryopreservation medium of the cryopreserved composition consists of dimethylsulfoxide at a concentration of 2% by weight, hydroxyethyl starch at a concentration of 6% by weight and water
Implementation Method 2
maintaining the composition at a temperature in the range of -60°C to -100°C, and wherein said maintaining comprises cooling the composition in a mechanical freezer
Data Source
AI summary
Disclosed are viable cell compositions and related methods of preparation, maintenance and use. The viable cell composition can contain specified levels of cells, hydroxy ethyl starch, and dimethylsulf oxide, and can be cryopreserved. The cryopreserved form of the composition can be thawed and combined with an aqueous liquid diluting medium to prepare a diluted viable cell composition that can contain specified, reduced levels of the dimethylsulfoxide and hydroxyethyl starch. The diluting medium can contain trehalose. The prepared, diluted viable cell composition can be administered to a patient.