Brown Adipocyte Stem Cell Amplification via Mechanical Dissociation

Resolve Bottlenecks,
Find Innovative SolutionsGenerate Solutions

Solution Overview

Problem

Current methods for in vitro or ex vivo amplification of brown or beige adipocytes from human adipose tissue are inefficient, particularly in obese patients, due to the rarity of these cells in adults and the destruction of three-dimensional tissue structures during standard enzymatic dissociation procedures, leading to contamination risks and low yields of therapeutic-grade cells.

Innovation Solution

A method involving mechanical dissociation to extract a stromal vascular fraction and extracellular matrix from adipose tissue, which are then cultured in suspension with a cell proliferation medium, allowing for three-dimensional amplification and reducing contamination risks, while maintaining the native tissue structure and endothelial cells.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Productivity

If enzymatic dissociation is used to isolate adipocyte precursors, then cell isolation efficiency is improved, but tissue structure is destroyed and contamination risk increases

Engineering Contradiction:

Improvecell isolation efficiencyVSAvoidtissue structure integrity

Core Design Contradiction:

ProductivityVSReliability

Solution Approach 1:

The patent replaces enzymatic dissociation (chemical system) with mechanical dissociation methods including mincing, squeezing through mesh, and agitation. This substitution maintains tissue structure integrity while achieving effective cell isolation, directly resolving the contradiction between isolation efficiency and tissue structure preservation.

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

Solution Approach 2:

The patent changes the dissociation parameter from enzymatic to mechanical force, and optimizes mesh size (e.g., 40-100 μm) and agitation intensity to achieve both effective cell separation and tissue structure maintenance. This parameter optimization resolves the contradiction by finding the right balance point.

Inventive Principle:

Principle #35Parameter changes

2Reliability

If non-enzymatic dissociation is used to reduce contamination, then contamination risk is reduced, but adipocyte precursor yield is low

Engineering Contradiction:

Improvecontamination reductionVSAvoidadipocyte precursor yield

Core Design Contradiction:

ReliabilityVSProductivity

Solution Approach 1:

The patent merges multiple mechanical dissociation steps (mincing, mesh filtration, agitation) into a combined protocol that achieves both low contamination and high yield. This merging of operations resolves the contradiction by creating a synergistic effect where each step contributes to both goals simultaneously.

Inventive Principle:

Principle #5Merging (Combining)

Solution Approach 2:

The patent performs preliminary mechanical dissociation and filtration steps before cell culture to remove contaminants and isolate pure adipocyte precursors. This preliminary action ensures both contamination reduction and high yield by preparing a clean cell population early in the process.

Inventive Principle:

Principle #10Preliminary action

3Productivity

If 2D culture is used for cell amplification, then cell expansion is achieved, but three-dimensional tissue structure is lost

Engineering Contradiction:

Improvecell amplificationVSAvoidthree-dimensional tissue structure

Core Design Contradiction:

ProductivityVSShape

Solution Approach 1:

The patent transitions from 2D culture to 3D suspension culture by forming cell aggregates in hanging drop or microfluidic devices. This dimensional change allows cell amplification while preserving three-dimensional tissue structure, directly resolving the contradiction between amplification and structure maintenance.

Inventive Principle:

Principle #17Another dimension (Dimensionality change)

Solution Approach 2:

The patent uses microfluidic devices with flexible chambers to form 3D cell aggregates in suspension. These flexible structures enable cell amplification in three dimensions while maintaining tissue-like architecture, resolving the contradiction between 2D expansion and 3D structure preservation.

Inventive Principle:

Principle #30Flexible shells and thin films

4Productivity

If standard enzymatic dissociation procedures are used, then cell isolation is achieved, but endothelial cells are lost

Engineering Contradiction:

Improvecell isolationVSAvoidendothelial cell loss

Core Design Contradiction:

ProductivityVSLoss of substance

Solution Approach 1:

The patent replaces enzymatic dissociation with gentle mechanical methods that do not damage endothelial cells. This substitution achieves cell isolation while preserving endothelial cell integrity and function, resolving the contradiction between isolation effectiveness and endothelial cell preservation.

Inventive Principle:

Principle #28Mechanics substitution (Replace mechanical system)

Data Source

PatentUS20230279355A1Method for the in vitro or ex vivo amplification of stem cells of brown or beige adipocytes

Publication Date: 2023.09.07 ESTAB FR DU SANG

AI summary

The method for the in vitro or ex vivo amplification of stem cells of brown or beige adipocytes includes: extracting (i) a stromal vascular fraction from human adipose tissue including endothelial cells of the vascular network of human adipose tissue and stem cells of brown or beige human adipose tissue and (ii) an extracellular matrix of the human adipose tissue, the extracellular matrix including endothelial cells of the vascular network of human adipose tissue, stem cells of brown or beige human adipose tissue and collagen; mixing the stromal vascular fraction and the extracellular matrix; and culturing the mixture obtained, in suspension, in a culture medium.