Stool Microorganism Viability via Iodixanol Cryoprotection

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Solution Overview

Problem

Current methods for storing stool microbiota for fecal transplantation face challenges due to the instability of microorganisms over time, particularly after antibiotic treatment, where the microbiota biodiversity is reduced, and finding a suitable donor for fecal transplantation can be difficult.

Innovation Solution

A method involving the separation of stool samples into fractions using density gradient centrifugation with iodixanol as a cryoprotectant, followed by freezing to preserve microorganisms in a viable form, allowing for long-term storage and subsequent reintroduction into a patient.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Loss of time

If stool samples are stored for a longer period of time for fecal transplantation, then donor availability and treatment flexibility are improved, but the microorganisms in the stool can change and viability is reduced

Engineering Contradiction:

Improvestorage timeVSAvoidmicroorganism viability

Core Design Contradiction:

Loss of timeVSReliability

Solution Approach 1:

The stool sample is separated into different density fractions using density gradient centrifugation, isolating the microorganism-containing fraction from other components. This segmentation allows selective preservation of viable microorganisms while removing substances that may degrade during storage.

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

Iodixanol serves as an intermediary substance with dual functionality: it creates the density gradient for separation and acts as a cryoprotectant during freezing. This intermediary protects microorganisms from ice crystal formation while enabling long-term storage.

Inventive Principle:

Principle #24Intermediary (Mediator)

2Manufacturing precision

If density gradient centrifugation is used to separate stool samples, then microorganism concentration and purity are improved, but processing complexity increases

Engineering Contradiction:

Improvemicroorganism separation purityVSAvoidprocessing steps

Core Design Contradiction:

Manufacturing precisionVSDevice complexity

Solution Approach 1:

Iodixanol performs multiple functions simultaneously: it acts as the density gradient medium for separation, serves as a cryoprotectant for preserving microorganism viability during freezing, and provides a stable storage environment. This multi-functionality reduces the need for additional separate processing steps.

Inventive Principle:

Principle #6Universality (Multi-functionality)

3Duration of action of stationary object

If stool samples are frozen for long-term storage, then storage duration is improved, but microorganism cell integrity may be compromised

Engineering Contradiction:

Improvestorage durationVSAvoidcell integrity

Core Design Contradiction:

Duration of action of stationary objectVSStability of the object's composition

Solution Approach 1:

Iodixanol is added to the microorganism fraction before freezing to provide cryoprotection. This beforehand cushioning prevents ice crystal formation that would otherwise damage cell membranes, ensuring microorganisms remain viable after prolonged storage and thawing.

Inventive Principle:

Principle #11Beforehand cushioning (Prior cushioning)

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 effectively maintains the viability of microorganisms during prolonged storage, ensuring their functionality and integrity for later use in fecal transplantation, addressing the issue of reduced biodiversity and donor availability.

Implementation Method 1

separating a stool sample into several fractions, forming a microorganism fraction which contains microorganisms from the intestinal microbiome

Methodology Applied

Scientific EffectDensity gradient centrifugation: Centrifugal Separation

Implementation Method 2

density gradient centrifugation with iodixanol

Methodology Applied

Scientific EffectDensity gradient: Density Gradient

Implementation Method 3

iodixanol as a cryoprotectant, followed by freezing to preserve microorganisms in a viable form

Methodology Applied

Scientific EffectCryoprotection: Freezing

Implementation Method 4

Freezing the microorganism fraction for preservation in such a way that the microorganisms are at least partially preserved in a viable form

Methodology Applied

Scientific EffectFreezing: Freezing

Data Source

PatentEP4273223A1Method for extracting and preserving microorganisms from a stool sample of a living being

Publication Date: 2023.11.08 LAB LS SE & CO KG

AI summary

Method for extracting and preserving microorganisms from a stool sample of a living being, in particular a human being, comprising the following steps: - Separating a stool sample into several fractions, resulting in a microorganism fraction containing microorganisms of the intestinal microbiome of the living being; - Freezing the microorganism fraction for preservation in such a way that the microorganisms are at least partially retained in a viable form.