Trehalose Cell-Washing Solution for Stem Cell Viability

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Solution Overview

Problem

Proteolytic enzyme treatments during stem cell subculture often result in cell damage and death, which is a significant challenge for providing high-quality stem cells for regenerative medicine applications.

Innovation Solution

Using a trehalose-containing physiological aqueous solution for cell washing before proteolytic enzyme treatment, specifically in concentrations of 0.1 to 20% w/v and treatment times of 1 second to 20 minutes, effectively suppresses cell death by inhibiting apoptosis and promoting cell damage repair pathways.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If proteolytic enzyme treatment is used to detach stem cells from culture vessels, then cell detachment efficiency is improved, but cell damage and death rate increases

Engineering Contradiction:

Improvecell detachment efficiencyVSAvoidcell viability

Core Design Contradiction:

ProductivityVSReliability

Solution Approach 1:

The patent applies preliminary action by washing cells with trehalose-containing solution before proteolytic enzyme treatment. This pre-treatment prepares the cells for subsequent enzyme exposure, reducing their susceptibility to damage. The trehalose is introduced in advance to protect cellular structures before the harmful enzyme action begins, thereby maintaining higher cell viability while still achieving effective detachment.

Inventive Principle:

Principle #10Preliminary action

Solution Approach 2:

Trehalose acts as an intermediary substance between the cells and the proteolytic enzyme. It mediates the interaction by forming a protective environment around the cells during enzyme treatment. The trehalose solution serves as a buffer that reduces the direct harmful impact of the enzyme on cellular proteins and membranes, allowing detachment to occur with minimal cell death.

Inventive Principle:

Principle #24Intermediary (Mediator)

2Ease of operation

If conventional washing solutions are used before proteolytic enzyme treatment, then treatment simplicity is maintained, but cell death rate remains high

Engineering Contradiction:

Improvetreatment simplicityVSAvoidcell survival rate

Core Design Contradiction:

Ease of operationVSReliability

Solution Approach 1:

The patent changes the chemical composition parameter of the washing solution by incorporating trehalose at specific concentrations (0.1-20% w/v). This parameter modification transforms a simple washing step into a protective pre-treatment. The trehalose concentration is optimized to provide maximum protection without interfering with subsequent enzyme activity, thereby improving cell survival while maintaining procedural simplicity.

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

This method significantly reduces cell death rates to less than 30%, providing a high-quality cell suspension for transplantation by maintaining a higher percentage of viable stem cells.

Implementation Method 1

trehalose is also used as an active ingredient of an organ-protecting solution in transplanting the organ because it has the properties of stabilizing cell membrane and suppressing cell damage

Methodology Applied

Scientific EffectCell membrane stabilization:

Implementation Method 2

the washing of adherent cells with a trehalose-containing physiological aqueous solution before detaching the cells from a culture vessel by a proteolytic enzyme such as trypsin can suppress cell death due to the proteolytic enzyme treatment

Methodology Applied

Scientific EffectApoptosis suppression:

Implementation Method 3

such an effect by trehalose was considered to be due to the suppression of an apoptosis pathway or the promotion of the function repairing the cell damage

Methodology Applied

Scientific EffectCell damage repair promotion:

Data Source

PatentEP2712920B1Method of washing adherent cell using trehalose-containing cell-washing solution

Publication Date: 2017.01.04 OTSUKA PHARMACEUTICAL FACTORY INC

AI summary

It is to provide the use of trehalose, its derivative or a salt thereof for reducing death of adherent cells due to proteolytic enzyme treatment; a method of washing an adherent cell, capable of effectively suppressing cell death due to proteolytic enzyme treatment for detaching the adherent cell from a culture vessel and subsequent cell treatment; a method of producing a cell suspension for transplantation using the cell-washing solution; and a kit comprising the cell-washing solution. Trehalose or its derivative or a salt thereof is added to a physiological aqueous solution to prepare a cell-washing solution containing trehalose or its derivative or a salt thereof as an active ingredient. The cell-washing solution can be used to wash an adherent cell before detaching the adherent cell from a culture vessel by proteolytic enzyme treatment to suppress (inhibit) cell death (apoptosis or the like) due to the proteolytic enzyme treatment. The concentration of trehalose applied to the cell-washing solution may be a concentration capable of suppressing (inhibiting) the cell death (apoptosis or the like) due to the proteolytic enzyme treatment, and is typically 0.1 to 20 (w/v)%, preferably 1 to 15 (w/v)%, more preferably 1.0 to 7.0 (w/v)%, still more preferably 3 to 5.0 (w/v)%.