Tangential Flow Filtration for Exosome and MSC Isolation

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Solution Overview

Problem

Current methods for obtaining exosomes and extracellular matrix components from mesenchymal stem cells are inefficient, often resulting in contaminated and diluted preparations due to the discarding of these components in supernatant fractions during centrifugation processes.

Innovation Solution

A method utilizing tangential flow filtration to isolate and concentrate mesenchymal stem cells and exosomes, avoiding ultra-centrifugation, which allows for the separation of exosomes and extracellular matrix components while maintaining their therapeutic potential without genomic modifications or stressful conditions.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Manufacturing precision

If centrifugation is used to prepare exosomes, then exosomes can be separated from cell culture, but the extracellular matrix components are discarded in supernatant fractions resulting in contamination and dilution

Engineering Contradiction:

Improvepurity of exosome preparationVSAvoidloss of extracellular matrix components

Core Design Contradiction:

Manufacturing precisionVSLoss of substance

Solution Approach 1:

The patent divides the cell culture into distinct fractions using tangential flow filtration, separating exosomes from extracellular matrix components rather than discarding the supernatant. This segmentation allows both fractions to be collected and processed independently, preventing loss of valuable extracellular matrix components while maintaining exosome purity.

Inventive Principle:

Principle #1Segmentation

Solution Approach 2:

Instead of discarding the supernatant fraction containing extracellular matrix components as in traditional centrifugation methods, the patent recovers these components by collecting the permeate fraction through tangential flow filtration. This recovery principle eliminates substance loss while maintaining the purity of the exosome retentate fraction.

Inventive Principle:

Principle #34Discarding and recovering

2Productivity

If ultra-centrifugation is used to concentrate exosomes, then therapeutic yield increases, but the process becomes complex and time-consuming

Engineering Contradiction:

Improvetherapeutic yield of exosomesVSAvoidcomplexity of filtration system

Core Design Contradiction:

ProductivityVSDevice complexity

Solution Approach 1:

The patent replaces the complex ultra-centrifugation mechanical system with a tangential flow filtration system that uses membrane separation based on size exclusion. This substitution maintains high productivity by concentrating exosomes effectively while simplifying the overall process through continuous flow operation rather than sequential centrifugation steps.

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

Solution Approach 2:

The patent changes the separation parameter from density-based centrifugation to size-based membrane filtration. By using membranes with specific pore sizes (e.g., 0.2-1.0 micrometers), the system achieves effective exosome concentration and separation without the complexity of ultra-centrifugation, maintaining productivity while reducing operational complexity.

Inventive Principle:

Principle #35Parameter changes

3Quantity of substance

If traditional centrifugation methods are used, then exosomes can be isolated, but the preparation is diluted and requires large volumes

Engineering Contradiction:

Improveconcentration of exosomesVSAvoidvolume of preparation required

Core Design Contradiction:

Quantity of substanceVSVolume of stationary object

Solution Approach 1:

The patent extracts exosomes from the cell culture medium using tangential flow filtration with membranes that retain exosomes while allowing culture medium to pass through as permeate. This extraction method concentrates exosomes in a reduced volume retentate fraction, achieving high concentration without requiring large preparation volumes.

Inventive Principle:

Principle #2Taking out (Extraction)

Solution Approach 2:

The patent employs porous membrane materials with controlled pore sizes (0.2-1.0 micrometers) in the tangential flow filtration system. These porous membranes selectively retain exosomes while permitting culture medium to pass through, thereby concentrating exosomes in the retentate and reducing the volume of final preparation required.

Inventive Principle:

Principle #31Porous materials

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 enables the production of homogeneous, concentrated exosome and MSC compositions with enhanced therapeutic yields, preserving the integrity and efficacy of the isolated components.

Implementation Method 1

filtering the MSC fraction through a first section of a tangential flow filtration system to obtain a retentate comprising the mesenchymal stem cells

Methodology Applied

Scientific EffectTangential flow filtration: Filter (physical)

Implementation Method 2

filtering the first permeate through a second section of the tangential flow filtration system to generate a second permeate comprising exosomes and at least one extracellular matrix component

Methodology Applied

Scientific EffectSize-based filtration: Filter (physical)

Data Source

PatentUS12123022B2Mesenchymal stem cell compositions and methods of making

Publication Date: 2024.10.22 VITTI LABS

AI summary

Disclosures herein are directed to first compositions comprising exosomes and extracellular matrix components and their use thereof. Also described are methods and kits wherein these first compositions are packaged and used with second compositions comprising mesenchymal stem cells. The first and second compositions are derived from the same tissue source using tangential flow filtration.