Cryopreservation Medium pH and Composition for Cell Viability
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Solution Overview
Problem
Current methods for cryopreserving cells for therapeutic purposes face challenges such as cellular lesions during thawing, instability of cryopreserved products, and toxicity from commonly used excipients, leading to limited long-term stability and viability issues.
Innovation Solution
A composition comprising a physiologically acceptable medium with saccharides, amino acids, DMSO or C3-C5 alkanediols, antioxidants, and cells, maintained at a pH between 7.0 and 8.5, which is used for cryopreservation by mixing with cells and then freezing, to create a stable and non-toxic product for long-term storage and easy administration.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If cells are cryopreserved using simple DMSO-based formulations, then freezing stability is improved, but long-term stability and cell viability deteriorate
Solution Approach 1:
The patent uses a composite cryopreservation formulation containing multiple components: DMSO (cryoprotectant), human albumin (stabilizing protein), coenzyme Q10 (antioxidant), and L-cysteine (amino acid with protective properties). This composite approach synergistically addresses both freezing stability and long-term viability, overcoming the limitations of simple DMSO-based formulations.
Solution Approach 2:
Human albumin acts as an intermediary substance that stabilizes cell membranes and proteins during freeze-thaw cycles. Coenzyme Q10 and L-cysteine serve as mediators that reduce oxidative stress and protect cellular structures, thereby maintaining cell functionality over extended periods after thawing.
2Reliability
If cells are preserved fresh in liquid medium, then cell viability is maintained short-term, but storage duration and flexibility deteriorate
Solution Approach 1:
The patent optimizes specific parameters of the cryopreservation medium including pH (adjusted to 7.4 using NaHCO3 buffer), osmolarity (adjusted to 300 mOsm/L using sucrose), and concentrations of protective agents (albumin 4%, coenzyme Q10 10 µM, L-cysteine 100 µM). These parameter optimizations ensure high cell viability after thawing while enabling long-term storage stability.
3Reliability
If commonly used excipients are used for cryopreservation, then freezing protection is improved, but toxicity to patients increases
Solution Approach 1:
The patent employs biocompatible, physiologically acceptable substances that are naturally present in the human body (albumin, coenzyme Q10, L-cysteine, amino acids, vitamins). These substances provide effective cryoprotection without requiring post-thaw washing steps, eliminating toxicity concerns while maintaining simplicity and clinical applicability.
4Object-affected harmful factors
If cells are washed before administration to remove toxic elements, then toxicity is reduced, but cell viability and productivity deteriorate
Solution Approach 1:
The cryopreservation formulation is designed to be self-contained and biocompatible, requiring no post-thaw washing or additional manipulations. The composition itself (using physiologically acceptable substances) ensures both toxicity reduction and high cell viability, eliminating the need for washing steps that would otherwise cause cell loss.
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 solution achieves high cell viability and functionality post-thawing, allowing for direct injection without washing, with cell viability maintained at 90% or higher for several hours after thawing, and stability for months, addressing the limitations of existing methods.
Implementation Method 1
it is also essential to use cryoprotectants to preserve cell integrity and functionality
Implementation Method 2
cooling at a slow rate allows an ordered crystallization of freezable water outside the cells
Implementation Method 3
the cells dehydrate, shrink, and the water exits the cell
Data Source
AI summary
The present invention relates to a composition comprising, in a physiologically acceptable medium:a) at least one saccharide,b) at least one amino acid,c) DMSO or at least one C3-C5 alkanediol,d) at least one antioxidant, ande) cells for therapeutic purposes,said composition having a pH between 7.0 and 8.5, preferably between 7.0 and 8.3. It also relates to a method for the cryopreservation of at least one sample of cells for therapeutic purposes, comprising the following steps:i) mixing the sample of cells for therapeutic purposes with ingredients a) to d) above and a physiologically acceptable medium, so as to obtain a composition having a pH between 7.0 and 8.5, preferably between 7.0 and 8.3, thenii) freezing the composition obtained in step i).