Stem Cell-Derived BBB Model for Stable Drug Permeability
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Solution Overview
Problem
Current in vitro human blood-brain barrier models lack stability and reproducibility, with previous models showing fluctuations in TEER values and limited longevity, making them unsuitable for reliable drug transport studies across the human BBB.
Innovation Solution
A method involving the differentiation of CD34+ hematopoietic stem cells into endothelial cells, followed by co-culture with pericytes to induce BBB properties, resulting in stable expression of BBB markers and transporters for at least 20 days, with high reproducibility across different laboratories and donors.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If primary human brain endothelial cells are used to establish in vitro BBB models, then the model reflects human-specific physiology and metabolism, but the model lacks stability and reproducibility due to constraints in obtaining human tissue and loss of hBEC phenotype during culture
Solution Approach 1:
The patent changes the cellular origin parameter from primary human brain endothelial cells to human pluripotent stem cells. This parameter change allows for unlimited proliferation while maintaining human-specific physiology, thereby resolving the contradiction between reliability and stability.
Solution Approach 2:
The patent performs preliminary differentiation of human pluripotent stem cells into brain endothelial cells under controlled conditions before establishing the BBB model. This preliminary action ensures that the cells are properly primed to exhibit stable, reproducible human BBB characteristics.
2Stability of the object's composition
If immortalized human brain endothelial cell lines are used, then the model provides long-term stability and reproducibility, but the model loses important tight junction properties and exhibits low TEER values
Solution Approach 1:
The patent performs preliminary differentiation of human pluripotent stem cells into brain endothelial cells under controlled conditions before establishing the BBB model. This preliminary action ensures that the cells are properly primed to exhibit stable, reproducible human BBB characteristics, including tight junction formation and appropriate TEER values.
Solution Approach 2:
The patent changes the cellular origin parameter from immortalized cell lines to freshly differentiated stem cell-derived endothelial cells. This parameter change maintains proliferative capacity while preserving tight junction functionality and TEER properties.
3Productivity
If iPSC-derived BBB models are used, then the model provides unlimited cell supply and human-specific characteristics, but the model shows low stability and high variability across different iPSC lines
Solution Approach 1:
The patent performs preliminary differentiation of human pluripotent stem cells into brain endothelial cells under controlled conditions before establishing the BBB model. This preliminary action ensures that the cells are properly primed to exhibit stable, reproducible human BBB characteristics, reducing variability across different iPSC lines.
Solution Approach 2:
The patent optimizes the differentiation protocol parameters to ensure consistent brain endothelial cell generation from various iPSC lines. This parameter optimization maintains high cell supply capacity while improving model stability and reproducibility.
Data Source
Figure 1A~1I
Figure 2A~2F
Figure 3A~3D
AI summary
The present disclosure relates to a method for obtaining human brain-like endothelial cells by contacting a population of cells isolated from stem cells with a differentiation medium to obtain endothelial cells and co- culturing said endothelial cells with pericytes, with cells of the neurovascular unit or with a pericytes conditioned medium, to obtain brain-like endothelial cells. The present disclosure also relates to the use of the brain-like endothelial cells as an in vitro model of human blood-brain barrier and a kit for measuring blood-brain barrier permeability of a substance, comprising in vitro human endothelial cells.