Stem Cell Cryopreservation Formulation for Oxidative Damage Control

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Solution Overview

Problem

Existing cryopreservation methods for stem cells using DMSO and FBS face issues such as immune responses, inconsistent ingredient ratios, and oxidative damage from peroxide radicals, necessitating a serum-free and effective cryoprotective agent to maintain cell viability.

Innovation Solution

A cryopreservation formulation containing glutathione, human serum albumin, and DMSO in a Hartmann solution is developed to minimize oxidative damage and improve cell viability during freezing and thawing.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If DMSO and FBS are used as cryoprotective agents, then cell protection during freezing is improved, but immune responses and inconsistent ingredient ratios occur

Engineering Contradiction:
Improvecell protection during freezingVSAvoidimmune responses and infection risk
Core Design Contradiction:
ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The invention extracts and removes the problematic FBS component from the cryoprotective agent formulation, replacing it with defined human serum albumin and antioxidant supplements. This extraction eliminates the immune response and infection risks associated with FBS while maintaining the essential cryoprotective functions.

Inventive Principle:
Principle #2Taking out (Extraction)

Solution Approach 2:

The invention changes the chemical composition parameters of the cryoprotective agent by replacing animal serum with purified human serum albumin and adding specific antioxidants (vitamin C, vitamin E, glutathione). This parameter change transforms the formulation from an undefined animal product to a precisely controlled, safe, and consistent composition.

Inventive Principle:
Principle #35Parameter changes

2Reliability

If peroxide radicals are present during cryopreservation, then oxidative damage to cells increases, but cell viability decreases

Engineering Contradiction:
Improvecell viability after thawingVSAvoidoxidative damage from peroxide radicals
Core Design Contradiction:
ReliabilityVSObject-generated harmful factors

Solution Approach 1:

The invention applies preliminary anti-action by incorporating antioxidants (vitamin C, vitamin E, glutathione) into the cryoprotective formulation before freezing. These antioxidants proactively neutralize peroxide radicals before they can cause oxidative damage to cell membranes, proteins, and DNA, thereby preserving cell viability throughout the cryopreservation process.

Inventive Principle:
Principle #9Preliminary anti-action

Solution Approach 2:

The invention converts the harmful effect of peroxide radicals into a beneficial outcome by using antioxidants to scavenge these radicals. The oxidative stress that would normally damage cells is transformed into a controlled process where antioxidants selectively neutralize the harmful radicals, protecting cellular components and improving post-thaw cell survival.

Inventive Principle:
Principle #22Blessing in disguise (Convert harm into benefit)

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 formulation significantly enhances cell survival rates and reduces cell clumping, maintaining high viability and therapeutic efficacy for stem cells during storage and transport.

Implementation Method 1

Peroxide radicals cause oxidative damage to cells, including lipid peroxidation, protein oxidation, and DNA damage. Accordingly, as a result of extensive efforts to develop a cryopreservation formulation for cells such as stem cells that can minimize the decrease in cell viability due to peroxide radicals during cryopreservation of cells

Methodology Applied
Scientific EffectAntioxidant activity: Oxidation

Implementation Method 2

Cryoprotective agents reduce physical and chemical cell damage during freezing and thawing, thereby improving cell viability after thawing. Typically, a cryoprotective agent (CPA) consisting of 10% (v/v) dimethyl sulfoxide (DMSO) and 90% (v/v) fetal bovine serum (FBS) is used for cell cryopreservation

Methodology Applied
Scientific EffectCryoprotection: Freezing

Implementation Method 3

A cryopreservation formulation containing glutathione, human serum albumin, and DMSO in a Hartmann solution is developed to minimize oxidative damage and improve cell viability during freezing and thawing

Methodology Applied
Scientific EffectColloidal protection: Colloid

Data Source

PatentEP4670502A1Formulation for cell cryopreservation
Publication Date: 2025.12.31 ENCELL CO LTD
  • EP4670502A1 patent drawingFigure 1
  • EP4670502A1 patent drawingFigure 2
  • EP4670502A1 patent drawingFigure 3A

AI summary

The present invention relates to a formulation for cell cryopreservation that can significantly improve the cell survival rate of cryopreserved cells, preferably stem cells. According to the present invention, the cell survival rate can be enhanced during the thawing process after freezing, thereby preventing the reduction in therapeutic efficacy that may occur during the storage and transportation of cell therapy products.