TGF-β and SHH Regulate Pluripotent Stem Cell Differentiation into Brain Microvascular Endothelial Cells in Generating...

Area of Science:

• Neuroscience
• Stem Cell Biology
• Biotechnology

Background:

• In vitro blood-brain barrier (BBB) models are crucial for central nervous system (CNS) research.
• Existing models using animal or cell line-derived cells have limitations.
• Generating mature brain microvascular endothelial-like cells (BME-like cells) from human pluripotent stem cells (hPSCs) for BBB modeling is challenging.

Purpose of the Study:

• To develop an efficient method for differentiating hPSCs into BME-like cells suitable for human BBB models.
• To create a reliable cellular platform for CNS drug delivery and disease research.

Main Methods:

• hPSCs were differentiated into BME-like cells using modulated TGF-β and SHH signaling pathways.
• Co-differentiation with neural cells mimicked in vivo BBB development.
• Barrier properties were assessed using transendothelial electrical resistance (TEER) and transporter activity assays.

Main Results:

• The differentiated BME-like cells exhibited well-organized tight junctions and appropriate transporter expression.
• These cells demonstrated polarized efflux transporter activity.
• BME-like cells responded to astrocytes, enhancing barrier function and showing immune quiescence by downregulating adhesion molecules.

Conclusions:

• A novel and efficient method for generating human BME-like cells from hPSCs was established.
• The developed BME-like cells form a functional human BBB model with desirable properties.
• This cellular platform holds promise for drug screening and developing brain-permeable pharmaceuticals.