High-Density Seeding of Embryoid Bodies for Stem Cell Purification

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Solution Overview

Problem

Current methods for differentiating pluripotent stem cells into desired cells, such as cardiomyocytes, face challenges in achieving a high proportion of desired cells while minimizing the presence of other cell types and cells with tumorigenic potential, resulting in low recovery rates and increased risks of tumorigenesis.

Innovation Solution

Seeding embryoid bodies at a density reaching confluence or higher to promote contact inhibition, thereby increasing the proportion of desired cells and reducing the proliferation of other cell types, and using treatments like Brentuximab-Vedotin to remove cells with tumorigenicity.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Quantity of substance

If conventional differentiation methods are used to generate desired cells from pluripotent stem cells, then desired cells can be obtained, but the proportion of desired cells in the cell population remains low and recovery rate is low

Engineering Contradiction:

Improveproportion of desired cellsVSAvoidrecovery rate

Core Design Contradiction:

Quantity of substanceVSProductivity

Solution Approach 1:

The invention changes the seeding density parameter to a very high level (confluence or higher) to exploit contact inhibition effects. This parameter change transforms the cell culture dynamics, causing non-desired cells to stop proliferating while desired cells continue to differentiate and survive, thereby increasing both the proportion and recovery rate of desired cells

Inventive Principle:

Principle #35Parameter changes

Solution Approach 2:

The invention converts the harmful effect of contact inhibition (which normally stops all cell proliferation) into a beneficial selective pressure. By seeding at confluence or higher, the contact inhibition selectively suppresses non-desired cells while desired cells, which are actively differentiating, can overcome this inhibition and continue to proliferate, thus turning a potentially harmful effect into a purification mechanism

Inventive Principle:

Principle #22Blessing in disguise (Convert harm into benefit)

2Quantity of substance

If conventional differentiation methods are used, then cell population can be generated, but cells with tumorigenic potential remain present

Engineering Contradiction:

Improvecell populationVSAvoidtumorigenic potential

Core Design Contradiction:

Quantity of substanceVSObject-affected harmful factors

Solution Approach 1:

The invention converts the harmful effect of contact inhibition into a beneficial selective pressure that eliminates tumorigenic cells. By seeding at confluence or higher, the contact inhibition selectively suppresses cells with abnormal proliferation characteristics (including tumorigenic cells) while allowing desired differentiating cells to continue their development, thereby reducing tumorigenic potential in the final cell population

Inventive Principle:

Principle #22Blessing in disguise (Convert harm into benefit)

3Quantity of substance

If seeding density is increased to confluence or higher, then proportion of desired cells increases, but resource consumption and culture complexity increase

Engineering Contradiction:

Improveproportion of desired cellsVSAvoidculture conditions

Core Design Contradiction:

Quantity of substanceVSDevice complexity

Solution Approach 1:

The invention employs a self-service mechanism where the high seeding density automatically creates contact inhibition conditions that perform the purification function. The system self-regulates by allowing desired cells to differentiate and proliferate while non-desired cells naturally stop dividing due to contact inhibition, eliminating the need for complex external control mechanisms or multiple purification steps

Inventive Principle:

Principle #25Self-service

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 increases the proportion of desired cells, such as cardiomyocytes, while decreasing the presence of cells with high proliferation rates and tumorigenic potential, resulting in a higher recovery rate of viable cells with reduced tumorigenic risk, and is compatible with conventional cell culture methods, making it suitable for producing sheet-shaped cell cultures.

Implementation Method 1

seeding a cell population by dispersing embryoid bodies obtained by inducing differentiation from the pluripotent stem cells to the desired cells, in which the embryoid bodies are dispersed at a density reaching confluence or higher... a density at which cell proliferation is substantially stopped by contact inhibition

Methodology Applied

Scientific EffectContact inhibition:

Data Source

PatentUS12084683B2Method for increasing the proportion of desired cells from induced pluripotent stem cells

Publication Date: 2024.09.10 TERUMO KK

AI summary

A method for producing a cell culture containing desired cells is provided. The desired cells are induced to be differentiated from pluripotent stem cells. The method includes seeding a cell population by dispersing embryoid bodies obtained by inducing differentiation from the pluripotent stem cells to the desired cells. The embryoid bodies are dispersed at a density reaching confluence or higher.