96 perfusable blood vessels to study vascular permeability in vitro

Area of Science:

• Vascular Biology
• Biomedical Engineering
• Cell Culture Technology

Background:

• Current in vitro vascular models use 2D monolayers, lacking physiological relevance.
• These models fail to replicate the tubular structure and microenvironment of native vasculature.
• There is a critical need for advanced in vitro models that mimic in vivo vascular architecture and function.

Purpose of the Study:

• To develop a robust, high-throughput 3D in vitro model of microvessels.
• To create a quantitative, real-time assay for assessing vascular barrier function.
• To enhance the physiological relevance of vascular research models.

Main Methods:

• Established a method to culture endothelial cells into 96 perfusable 3D microvessels.
• Optimized culture conditions for microvessel formation within 7 days, with viability exceeding 60 days.
• Developed a real-time assay to measure microvessel permeability.

Main Results:

• The 3D microvessels demonstrated selective permeability, allowing passage of 20 kDa dextran but not 150 kDa dextran.
• Vascular endothelial growth factor (VEGF) and tumor necrosis factor-alpha (TNFα) elicited dose-dependent responses, confirming physiological relevance.
• The model system proved robust and suitable for high-throughput screening.

Conclusions:

• The developed 3D microvessel culture and permeability assay provide a more accurate in vitro model for vascular studies.
• This method facilitates the transition from 2D to 3D culture systems in vascular biology.
• The platform enables advanced research into vascular barrier function and responses to stimuli.