hPSC-Derived Brain Endothelial Cells for BBB Model Fidelity

Solution Overview

Problem

Current in vitro blood-brain barrier (BBB) models, particularly those of human origin, face challenges due to limitations in availability and fidelity, with previous models exhibiting poor barrier properties and species differences, hindering high-throughput screening and studies on brain-penetrating molecules and BBB developmental pathways.

Innovation Solution

A method is developed to produce human pluripotent stem cell (hPSC)-derived brain-specific endothelial cells by inducing differentiation in unconditioned medium without fibroblast growth factor, followed by expansion in endothelial cell medium, and co-culturing with astrocytes or other cell types, resulting in cells expressing markers like GLUT-1, PECAM-1, claudin-5, and p-glycoprotein, and achieving high trans-endothelial electrical resistance.

Engineering Contradictions & Design Principles

VSEngineering Contradiction Analysis

1Adaptability or versatility

If primary human BMECs are isolated from autopsy tissue or resected brain specimens, then human origin is achieved, but availability and fidelity are limited

Engineering Contradiction:

Improvehuman originVSAvoidavailability

Core Design Contradiction:

Adaptability or versatilityVSEase of manufacture

Solution Approach 1:

The patent uses pluripotent stem cells (PSCs) as a renewable source to generate human BMECs in vitro, creating a copy of the target cell type that can be produced on demand. This eliminates the need to isolate cells from limited autopsy or surgical specimens, thereby improving availability while maintaining human origin and fidelity.

Inventive Principle:

Principle #26Copying

2Ease of manufacture

If immortalized BMECs are used, then availability is improved, but barrier properties deteriorate with low TEER and discontinuous tight junction expression

Engineering Contradiction:

ImproveavailabilityVSAvoidbarrier properties

Core Design Contradiction:

Ease of manufactureVSReliability

Solution Approach 1:

The patent employs small molecule compounds that specifically modulate key signaling pathways (such as Wnt/β-catenin, Notch, and VEGF pathways) to induce and maintain high barrier properties in PSC-derived BMECs. This allows the cells to achieve and sustain high TEER values and continuous tight junction expression, overcoming the barrier property deterioration seen in immortalized cells while maintaining availability.

Inventive Principle:

Principle #35Parameter changes

3Measurement precision

If species differences are avoided by using human cells, then fidelity is improved, but model robustness and scalability are reduced

Engineering Contradiction:

ImprovefidelityVSAvoidscalability

Core Design Contradiction:

Measurement precisionVSProductivity

Solution Approach 1:

The patent uses PSCs as a renewable source to generate human BMECs in vitro, creating a copy of the target cell type that can be produced on demand. This eliminates the need to isolate cells from limited autopsy or surgical specimens, thereby improving availability while maintaining human origin and fidelity.

Inventive Principle:

Principle #26Copying

Solution Approach 2:

The patent employs small molecule compounds that specifically modulate key signaling pathways (such as Wnt/β-catenin, Notch, and VEGF pathways) to induce and maintain high barrier properties in PSC-derived BMECs. This allows the cells to achieve and sustain high TEER values and continuous tight junction expression, overcoming the barrier property deterioration seen in immortalized cells while maintaining availability.

Inventive Principle:

Principle #35Parameter changes

Data Source

PatentUS9902940B2Human blood-brain barrier endothelial cells derived from pluripotent stem cells and blood-brain barrier model thereof

Publication Date: 2018.02.27 WISCONSIN ALUMNI RES FOUND

AI summary

A model blood brain barrier obtained from hPSCs is disclosed.