Non-Toxic Cryoprotective Composition for Cellular Integrity

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Solution Overview

Problem

Traditional cryopreservation techniques cause cellular damage due to ice crystal formation and osmotic issues, leading to cell death during freezing and thawing, and existing cryoprotectants like DMSO and glycerol are toxic and cause side effects.

Innovation Solution

A composition comprising polyethylene glycol (PEG), saccharides such as trehalose and dextran-40, and organic amphoteric agents like ectoin and hydroxyectoin is used to protect cells and tissues during cryopreservation, maintaining cellular integrity and viability through multiple freeze/thaw cycles.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Reliability

If traditional cryoprotectants like DMSO and glycerol are used, then cellular protection during freezing is achieved, but toxicity and side effects occur

Engineering Contradiction:
Improvecellular protectionVSAvoidtoxicity
Core Design Contradiction:
ReliabilityVSObject-affected harmful factors

Solution Approach 1:

The patent changes the chemical parameters of cryoprotective agents by using non-toxic alternatives such as sugars (trehalose, sucrose), polyols (sorbitol, mannitol), and amino acids (taurine, glycine) instead of traditional toxic cryoprotectants like DMSO and glycerol. These alternative substances provide comparable or superior cellular protection without the harmful side effects, thereby resolving the contradiction between effectiveness and toxicity.

Inventive Principle:
Principle #35Parameter changes

Solution Approach 2:

The patent employs naturally occurring, biocompatible substances that are non-toxic and can be easily removed or metabolized by cells. These include common sugars and amino acids that are already present in biological systems, making them safe for prolonged cell storage and eliminating the need for complex removal procedures required by toxic cryoprotectants.

Inventive Principle:
Principle #27Cheap short-living objects (Disposable)

2Duration of action of stationary object

If freezing is performed to preserve cells, then biological activity is stopped, but ice crystal formation causes cellular damage

Engineering Contradiction:
Improvepreservation timeVSAvoidice crystal damage
Core Design Contradiction:
Duration of action of stationary objectVSObject-affected harmful factors

Solution Approach 1:

The patent introduces intermediary substances (cryoprotective agents such as trehalose, sucrose, sorbitol, mannitol, taurine, and glycine) that mediate between the freezing process and cellular structures. These intermediaries prevent direct contact between ice crystals and cell membranes, thereby protecting cells from mechanical damage while allowing long-term frozen storage.

Inventive Principle:
Principle #24Intermediary (Mediator)

Solution Approach 2:

The patent applies cryoprotective agents to cells before freezing to provide preemptive protection. These agents accumulate around cellular structures in advance, creating a protective environment that cushions cells against the formation and expansion of ice crystals during the freezing process, thereby preventing cellular damage before it occurs.

Inventive Principle:
Principle #11Beforehand cushioning (Prior cushioning)

3Adaptability or versatility

If freezing and thawing cycles are performed, then cell storage and retrieval is enabled, but osmotic stress leads to cell death

Engineering Contradiction:
Improvestorage and retrieval capabilityVSAvoidcell viability
Core Design Contradiction:
Adaptability or versatilityVSReliability

Solution Approach 1:

The patent modifies the osmotic parameters of the freezing medium by incorporating non-toxic cryoprotective agents that maintain osmotic balance during freeze-thaw cycles. Substances like sugars and polyols adjust the osmotic pressure to prevent excessive water movement in and out of cells, thereby maintaining cell viability through multiple storage and retrieval cycles.

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 composition effectively prevents cellular damage by reducing ice crystal formation and osmotic stress, improving post-thaw cell viability and function, and is non-toxic, thus overcoming the limitations of traditional cryoprotectants.

Implementation Method 1

Cryopreservation is a process where biological samples such as cells or whole tissues are preserved by cooling to low sub-zero temperatures

Methodology Applied
Scientific EffectCryoprotection: Freezing

Implementation Method 2

Traditional cryopreservation techniques cause cellular damage due to ice crystal formation and osmotic issues

Methodology Applied
Scientific EffectOsmotic pressure regulation: Osmotic Pressure

Data Source

PatentUS11985968B2Cryosolutions and uses thereof
Publication Date: 2024.05.21 AKRON BIOTECHNOLOGY LLC

AI summary

Compositions for maintaining cellular integrity especially in cases of cryopreservation and multiple freeze/thawing cycles are provided. In addition, the compositions can be used in various media for a variety of applications.