Lyophilized Cell Production via Freeze-Drying
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Solution Overview
Problem
Current cell preservation methods, such as cryopreservation, face challenges including osmotic stress, cell viability inconsistencies, and logistical issues during storage and transport, particularly due to the use of cryoprotectants like DMSO, which can be toxic and limit the effectiveness of sugars as alternatives.
Innovation Solution
A method involving the lyophilization of cells by freezing a composition containing cells, a polyol, a sugar, and a polysaccharide, followed by the removal of at least 90% of the aqueous component, allowing for the production of stable lyophilized cells that can remain viable upon reconstitution, thereby avoiding the limitations of traditional cryopreservation.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If cryopreservation is used to store cells, then cell viability can be maintained, but the process requires toxic cryoprotectants like DMSO and complex freezing procedures that cause osmotic stress and cell shrinkage
Solution Approach 1:
The patent changes the physical state parameter of the cells from frozen (cryopreservation) to lyophilized (freeze-dried). This parameter change allows cells to be preserved without requiring toxic cryoprotectants like DMSO, and eliminates the osmotic stress and cell shrinkage associated with freezing. The lyophilization process maintains cell viability while removing the harmful factors inherent in cryopreservation.
Solution Approach 2:
The patent extracts and removes the aqueous component from the cell composition through lyophilization. By removing water and ice formation, the process eliminates the source of osmotic stress that causes cell dehydration and shrinkage during freezing. This extraction of the problematic aqueous phase allows cell preservation without the harmful effects of cryopreservation.
2Stability of the object's composition
If slow cooling is used during freezing, then ice crystal formation is reduced, but hypertonic solution is generated causing cell dehydration and shrinkage
Solution Approach 1:
The patent utilizes phase transition from frozen to lyophilized state. By freezing the cells and then removing the ice through sublimation under vacuum, the process avoids the hypertonic solution generation that occurs during slow cooling. The phase change from solid ice to vapor bypasses the liquid phase where osmotic stress occurs, preserving both cell structure and volume.
3Object-affected harmful factors
If rapid cooling is used during freezing, then osmotic stress is avoided, but intracellular ice crystals are formed causing cell injury
Solution Approach 1:
The patent introduces lyophilization as an intermediary process between freezing and cell storage/transport. The freeze-drying process removes ice crystals through sublimation under vacuum, eliminating the intracellular ice formation that causes cell injury during rapid cooling. This intermediary step preserves cell viability while avoiding both osmotic stress and ice crystal damage.
4Reliability
If cryopreservation is used for long-term storage and transport, then cell viability is maintained, but logistical complexity and storage requirements increase
Solution Approach 1:
The patent changes the storage parameter from frozen (requiring -80°C or liquid nitrogen) to lyophilized (stable at refrigerated or ambient temperatures). This parameter change dramatically simplifies logistics by eliminating the need for complex cryogenic storage infrastructure, specialized transport containers, and continuous monitoring systems, while maintaining cell viability through the stable lyophilized state.
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 enables the production of viable cells that can be stored at increased temperatures, reducing the need for cryoprotectants and improving the stability and consistency of cell viability during storage and transport.
Implementation Method 1
freezing a composition comprising a population of cells, an aqueous component, a polyol, a sugar, and a polysaccharide
Implementation Method 2
removing at least 90% of the aqueous component from the frozen composition to produce the population of lyophilized cells
Data Source
AI summary
The present disclosure provides a method of producing a population of lyophilized cells, comprising: (a) freezing a composition comprising a population of cells, an aqueous component, a polyol, a sugar, and a polysaccharide; and (b) removing at least 90% of the aqueous component from the frozen composition to produce the population of lyophilized cells. On some embodiments, the disclosure provides a method of producing a population of reconstituted viable cells, comprising: (a) freezing a composition comprising a population of cells, an aqueous component, a polyol, a sugar, and a polysaccharide; (b) removing at least 90% of the aqueous component from the frozen composition to produce the population of lyophilized cells, and (c) resuspending the population of lyophilized cells in a reconstitution agent to form a reconstituted composition, wherein at least 1% of the cells are viable.