Urea Monosaccharide Cryoprotectant Composition for Cell Viability
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Solution Overview
Problem
Current methods for storing human mesenchymal stem cells (hMSCs) using cryopreservation in liquid nitrogen are cumbersome, expensive, and involve toxic cryoprotectants like DMSO and immunogenic fetal bovine serum, posing safety risks and requiring specialized handling, while alternative lyophilization methods are not effectively viable.
Innovation Solution
A composition comprising viable cells, an aqueous component, urea, and a monosaccharide, with optional bulking agents and hyaluronan gel, that allows for cryopreservation or lyophilization at controlled temperatures without DMSO, maintaining cell viability through specific concentration ranges and molar ratios of urea and monosaccharides.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If cryopreservation in liquid nitrogen is used to store hMSCs, then cell viability is maintained during long-term storage, but the process becomes cumbersome, expensive, and requires toxic additives like DMSO and FBS
Solution Approach 1:
The invention extracts and eliminates the problematic components DMSO and FBS from the cryopreservation system, replacing them with a simplified composition containing only urea, monosaccharide, and optional bulking agents. This extraction removes the toxicity and immunogenicity while maintaining the core function of cell preservation during freezing and lyophilization
Solution Approach 2:
The invention changes the chemical parameters of the cryoprotective composition by substituting traditional agents (DMSO, FBS) with alternative substances (urea at 0.2-1.25 M, monosaccharide at 0.2-1.25 M). This parameter change maintains cell viability while eliminating the need for complex handling procedures and specialized facilities
2Reliability
If DMSO and FBS are added to preserve cells during cryopreservation, then cell viability is maintained, but safety risks increase due to toxicity and immunogenicity
Solution Approach 1:
The invention converts the harmful effects of traditional cryoprotectants into beneficial properties by using urea and monosaccharides that are non-toxic and non-immunogenic. These substances naturally protect cells during freezing and lyophilization without introducing safety risks, thereby eliminating the need for post-storage removal procedures
Solution Approach 2:
The invention replaces expensive, hazardous materials (DMSO, FBS) with inexpensive, safe alternatives (urea, monosaccharides). This substitution eliminates safety risks while reducing costs, and the composition can be used without requiring specialized removal facilities since the alternatives are inherently safe
3Object-affected harmful factors
If specialized facilities and trained personnel are used to remove DMSO and FBS, then safety is improved, but the process becomes labor-intensive and complex
Solution Approach 1:
The invention extracts the need for specialized removal procedures by eliminating DMSO and FBS from the composition entirely. The replacement substances (urea, monosaccharides) are inherently safe and do not require removal, thereby simplifying the operational process and eliminating the need for specialized facilities and trained personnel
Solution Approach 2:
The invention enables the composition to be self-sufficient by using substances that are inherently safe and do not require external removal processes. The urea and monosaccharide composition automatically provides protection without introducing hazards that would need to be managed by specialized personnel
4Ease of operation
If lyophilization is used to store cells at refrigerated temperatures, then handling and shipping become easier, but traditional lyophilization methods do not maintain cell viability
Solution Approach 1:
The invention changes the chemical parameters of the lyophilization process by using a composition containing urea (0.2-1.25 M) and monosaccharide (0.2-1.25 M) instead of traditional protectants. This parameter change enables cells to survive the lyophilization process and remain viable at refrigerated temperatures, combining handling ease with cell viability
Solution Approach 2:
The invention uses a composite composition of urea and monosaccharide (with optional bulking agents) that provides both cryoprotective and lyoprotective properties. This composite material enables cells to withstand both freezing and lyophilization processes, making the cells suitable for storage at refrigerated temperatures while maintaining viability
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 viable cell storage at controlled temperatures, reducing the need for expensive cryogenic storage and toxic additives, while maintaining cell viability and safety, facilitating easier handling and shipping of hMSCs.
Implementation Method 1
The present disclosure provides for methods of providing cryoprotection and/or lyoprotection of cells that are stored by freezing or lyophilization
Implementation Method 2
lyophilize (freeze-dry) the cells
Implementation Method 3
removing at least 90% of the aqueous component from the frozen composition to produce the population of lyophilized cells
Data Source
AI summary
Provided are compositions comprising a population of cells that are amenable to freezing or lyophilization while maintaining viable cells. The compositions comprise a monosaccharide, urea and, in some embodiments, a bulking agent. Also provided are methods for preparing and storing cells via freezing or lyophilization in a manner that allows cells to remain viable during storage.