Cryoprotective Formulation for Cell Viability and Direct Injection
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Solution Overview
Problem
Current methods for cryopreserving cells for therapeutic purposes face challenges such as cell damage during thawing, instability of cryoprotectants, and toxicity of excipients, leading to reduced viability and flexibility in storage and administration of cell therapy products.
Innovation Solution
A composition comprising human serum albumin, a saccharide (preferably trehalose), DMSO, and L-cysteine or coenzyme Q10 is used to cryopreserve cells, which are then frozen at controlled temperatures, allowing for stable long-term storage and direct injection without washing, thereby reducing toxicity and handling-related cell loss.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If cells are cryopreserved using conventional methods with simple DMSO-based formulations, then freezing is achieved, but cell damage occurs during thawing leading to reduced viability
Solution Approach 1:
The patent uses a composite cryoprotective formulation containing human serum albumin, saccharides (such as trehalose), DMSO, and antioxidants (such as L-cysteine or coenzyme Q10). This composite material provides multiple protective functions simultaneously: albumin and saccharides prevent ice crystal formation and maintain osmotic balance, DMSO acts as a penetrating cryoprotectant, and antioxidants reduce oxidative stress during freezing and thawing. The synergistic combination resolves the contradiction by protecting cells from multiple damage mechanisms rather than relying on DMSO alone.
Solution Approach 2:
The formulation includes antioxidants (L-cysteine or coenzyme Q10) that are added before freezing to cushion against oxidative damage that occurs during the freezing and thawing process. The saccharides and albumin also prepare the cellular environment in advance by maintaining osmotic balance and preventing ice crystal formation, thereby cushioning cells against mechanical damage before the actual thawing occurs.
2Duration of action of stationary object
If cells are stored fresh in liquid medium with limited preservation, then cell functionality is maintained short-term, but storage duration is limited to a few hours or days
Solution Approach 1:
The patent changes the physical and chemical parameters of the storage medium by incorporating specific concentrations of cryoprotectants (DMSO at 2-10%, saccharides at 0.1-1 M), albumin (1-10% w/v), and antioxidants. These parameter changes enable the medium to support cell survival at ultra-low temperatures for extended periods while maintaining cell functionality upon thawing, thereby resolving the contradiction between storage duration and cell functionality.
3Object-affected harmful factors
If cells are washed before administration to remove toxic excipients, then patient toxicity is reduced, but additional handling operations cause decreased viability and cell loss
Solution Approach 1:
The patent employs a formulation designed for single-use direct injection without requiring washing steps. The cryoprotective formulation is optimized to be non-toxic at the concentrations used, eliminating the need for washing. This disposable approach resolves the contradiction by making the excipients inherently safe rather than requiring removal, thereby avoiding handling operations that would reduce cell viability.
4Duration of action of stationary object
If conventional cryopreservation formulations are used, then freezing is achieved, but the formulations are not stable and not effective in the long term
Solution Approach 1:
The patent creates a stable long-term formulation by combining multiple components with complementary stabilizing functions. Human serum albumin provides structural stability and protein protection, saccharides prevent ice crystal formation and maintain osmotic balance, DMSO acts as a penetrating cryoprotectant, and antioxidants prevent oxidative degradation. This composite material achieves both formulation stability and long-term effectiveness that simple DMSO-based formulations cannot provide.
Solution Approach 2:
The formulation includes human serum albumin and saccharides as intermediary substances that mediate between the cryoprotectants and the cells. These intermediaries stabilize the cellular environment during freezing and storage, preventing direct harmful interactions between DMSO and cellular components, thereby achieving long-term stability and effectiveness.
Data Source
AI summary
Disclosed is a composition including, in a physiologically acceptable medium: a) human albumin; b) at least one saccharide; c) DMSO and L-cysteine or coenzyme Q10; and d) cells for therapeutic purposes, with the exception of specific tumour-infiltrating lymphocytes.