DMSO-Free Cryopreservation Composition for Ice Crystal Control

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Solution Overview

Problem

Current cryopreservation methods using dimethyl sulfoxide (DMSO) are toxic and ineffective in controlling ice crystal growth and recrystallization during thawing, leading to low survival rates and safety concerns for cryopreserved cells and tissues.

Innovation Solution

A DMSO-free cryopreservation solution comprising biomimetic ice growth inhibition materials like polyvinyl alcohol (PVA) and amino acids, along with polyalcohols and water-soluble saccharides, to inhibit ice growth and enhance cell survival.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If DMSO is used as the main protective agent in cryopreservation reagents, then cell protection during freezing is improved, but cell toxicity and side effects increase

Engineering Contradiction:

Improvecell protection capabilityVSAvoidcell toxicity

Core Design Contradiction:

ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The invention extracts and removes DMSO from the cryopreservation reagent formulation, replacing it with alternative protective agents that do not exhibit the same toxic side effects while maintaining or improving cell protection capability during the freezing and thawing processes

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 2:

The invention changes the chemical composition parameters of the cryopreservation reagent by substituting DMSO with alternative protective agents, thereby altering the toxicity profile while preserving the essential freezing protection function

Inventive Principle:

Principle #35Parameter changes

2Reliability

If high concentration DMSO (≥15%) is used for vitrification, then freezing protection is improved, but survival rate and safety of cryopreserved objects deteriorate

Engineering Contradiction:

Improvefreezing protectionVSAvoidsurvival rate

Core Design Contradiction:

ReliabilityVSManufacturing precision

Solution Approach 1:

The invention removes high concentration DMSO from the vitrification solution and replaces it with alternative protective agents that achieve effective freezing protection without the severe toxic effects associated with high DMSO concentrations, thereby improving survival rates

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 2:

The invention employs a composite formulation combining multiple protective agents including amino acids, peptides, and other non-toxic compounds to achieve the freezing protection previously provided by high concentration DMSO, without the associated toxicity and survival rate deterioration

Inventive Principle:

Principle #40Composite materials

3Duration of action of stationary object

If current cryopreservation reagents are used, then freezing capability is maintained, but capability to control ice crystal growth and recrystallization during thawing is lost

Engineering Contradiction:

Improvefreezing capabilityVSAvoidice crystal control capability

Core Design Contradiction:

Duration of action of stationary objectVSReliability

Solution Approach 1:

The invention modifies the chemical composition parameters of the cryopreservation reagent by incorporating specific amino acids and peptides that possess ice crystal growth inhibition properties, thereby enhancing the capability to control ice crystal formation and recrystallization during both freezing and thawing processes while maintaining freezing protection

Inventive Principle:

Principle #35Parameter changes

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 and tissue survival rates comparable to or exceeding commercial DMSO-based methods, while eliminating toxicity and serum-related contaminants, ensuring stable function expression and preservation efficiency.

Implementation Method 1

the biomimetic ice growth inhibition material is selected from polyvinyl alcohol (PVA) and/or an amino acid biomimetic ice growth inhibition material

Methodology Applied

Scientific EffectIce growth inhibition:

Implementation Method 2

a DMSO-free cryopreservation solution comprising, per 100 mL in volume, 0.01-50.0 g of a biomimetic ice growth inhibition material, 5.0-45 mL of a polyalcohol, a water-soluble saccharide at 0.1-1 mol L−1

Methodology Applied

Scientific EffectCryopreservation:

Implementation Method 3

the vitrification technology can allow liquid inside and outside a cell to be directly vitrified in the rapid freezing process so as to avoid the damage resulting from the formation of ice crystals

Methodology Applied

Scientific EffectVitrification:

Data Source

PatentUS12408659B2DMSO-free cryopreservation solution and preparation method and use thereof

Publication Date: 2025.09.09 PEKING UNIVERSITY THIRD HOSPITAL (THE THIRD CLINICAL MEDICAL SCHOOL OF PEKING UNIVERSITY)

AI summary

A cryopreservation solution contains 0.01-50.0 g of bionic ice control materials, 5.0-30 mL of polyols, 1-30 g of water-soluble sugar, and 0-30 mL of serum, and a buffer in every 100 mL of the cryopreservation solution. It does not contain DMSO. When being used for the cryopreservation of mouse oocytes and embryos, the solution may achieve the same or an even higher cell and tissue survival rate and functional expression stability as or than a commercial cryopreservation solution (containing 15% DMSO), and has high preservation efficiency. The cryopreservation solution without DMSO or serum reduces parasitic biological contaminants in the commercial cryopreservation solution containing serum currently used in clinical practice.