Stem Cell Cryopreservation Using N-Acetylcysteine

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Solution Overview

Problem

Current methods for stem cell cryopreservation often result in a decline in cell viability and functional capabilities post-thaw, due to stresses from the freezing and thawing process, such as oxidative stress and membrane disruptions, leading to reduced cellular activity and reproductive ability.

Innovation Solution

Treatment of stem cells with N-acetylcysteine (NAC) prior to freezing and potentially after thawing, to enhance post-thaw viability, growth rate, and mitochondrial activity, while maintaining structural and functional properties.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If conventional cryopreservation methods using DMSO, glycerol or animal-derived serum are used, then cell protection during freezing is provided, but post-thaw cell viability and functional capabilities decline

Engineering Contradiction:

Improvecell viability post-thawVSAvoidoxidative stress and membrane disruptions

Core Design Contradiction:

ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The patent changes the chemical composition parameter of the cryopreservation medium by replacing conventional cryoprotectants (DMSO, glycerol, animal-derived serum) with a plant-based composition containing polyethylene glycol (PEG), sugars, and amino acids. This parameter change resolves the contradiction by providing cell protection during freezing while eliminating the harmful oxidative stress and membrane disruptions caused by conventional agents, thereby maintaining high post-thaw cell viability and functional capabilities

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The patent employs a disposable, plant-based cryopreservation composition that is metabolically benign and does not require long-term cell exposure. The composition is designed for short-term use during the freezing and thawing process, after which cells can be cultured in standard media. This approach resolves the contradiction by providing effective protection during the critical freezing/thawing window without the cumulative toxic effects of conventional cryoprotectants

Inventive Principle:

Principle #27Cheap short-living objects (Disposable)

2Duration of action of stationary object

If storage intervals are extended beyond 1-3 days for hypothermic preservation, then longer-term cell availability is achieved, but cell viability and function deteriorate

Engineering Contradiction:

Improvestorage intervalVSAvoidcellular responsiveness and function

Core Design Contradiction:

Duration of action of stationary objectVSReliability

Solution Approach 1:

The patent changes the physical state parameter by transitioning from hypothermic preservation (refrigerated storage at 4°C) to cryopreservation (freezing at -80°C or lower). This parameter change enables extended storage intervals while maintaining cell viability and function, as the frozen state halts metabolic activity and prevents the deterioration that occurs during prolonged hypothermic storage

Inventive Principle:

Principle #35Parameter changes

3Duration of action of stationary object

If the freezing and thawing process is performed, then cell preservation for later use is achieved, but cellular activity and reproductive ability are reduced

Engineering Contradiction:

Improvepreservation durationVSAvoidcellular activity and proliferation potential

Core Design Contradiction:

Duration of action of stationary objectVSProductivity

Solution Approach 1:

The patent applies preliminary action by pre-treating cells with the plant-based cryopreservation composition before freezing, which prepares cells for the stress of freezing and thawing. The composition is applied in advance to allow cells to adapt and accumulate protective mechanisms, thereby resolving the contradiction by enabling long-term preservation while maintaining high cellular activity and proliferation potential after thawing

Inventive Principle:

Principle #10Preliminary action

Solution Approach 2:

The patent converts the harmful effects of freezing and thawing (oxidative stress, membrane disruptions) into beneficial outcomes by using a composition that actively counteracts these stresses. The plant-based ingredients scavenger free radicals and stabilize membranes during the freezing/thawing process, thereby transforming what would be damaging events into a controlled process that preserves rather than deteriorates cellular function

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 use of NAC in stem cell cryopreservation methods increases the number of viable cells post-thaw, accelerates recovery, and improves growth rates, maintaining the cells' therapeutic potential and functionality.

Implementation Method 1

the production of free radicals by disruption of oxidative respiration, which are detrimental to cells due to the downstream effects of lipid peroxidation

Methodology Applied

Scientific EffectFree radical scavenging: Oxidation

Data Source

PatentUS20220104481A1Cryopreservation of stem cells

Publication Date: 2022.04.07 TAKEDA PHARMA CO LTD

AI summary

The invention relates to methods for the cryopreservation of a stem cell population, including mesenchymal stem cells (MSCs) such as adipose-derived stromal stem cells (ASCs). More particularly, the invention relates to the use of N-acetylcysteine (NAC) in cryopreservation methods, populations of cells obtained from said methods, compositions comprising said cells and uses thereof.