Blood Brain Barrier Model in a 3D Co-Culture Microfluidic System

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Images showing the immunocytochemistry of primary neurons, primary astrocyte and endothelial cells with specific cell type markers. A) Representative images showing top and side views of the three cell types in 3D co-culture. B) 3D view of of neuron channel. C) Representative images showing immature neurons identified by DCX, astrocytes characterized by GFAP, and HUVEC expressing VE-cadherin. The scale bars in A and B are 200um, and the scale bar in C is 50um.Images showing the endothelial barrier characterization. A&B) Representative image showing HUVEC (A) and  hCMEC/D3 (B)  monolayers expressing F-actin and VE-cadherin.  C&D) Three-dimensional visualizations. E&F) Sections of the endothelial walls for HUVEC (C&E) and hCMEC/D3 (D&F). Scale bar 50um.
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Inventors
Professor Roger Kamm
Department of Mechanical Engineering, MIT
External Link (web.mit.edu)
Giulia Adriani
Singapore-MIT Alliance For Research and Technology
Andrea Pavesi
Singapore-MIT Alliance For Research and Technology
Dong Liang Ma
National University of Singapore
Eyleen Lay Keow Goh
National University of Singapore
Managed By
Michelle Hunt
MIT Technology Licensing Officer
Patent Protection

Blood Brain Barrier Model in a 3D Co-culture Microfluidic System

PCT Patent Application WO 2017-035119

Applications

This invention is a blood-brain barrier model that may be used in drug delivery studies.

Problem Addressed

The Blood-brain barrier (BBB) is a selective and distributed barrier implemented by tight junctions between endothelial cells forming the vascular walls of capillaries in the central nervous system that protects the system from damage. Unfortunately, the BBB also hampers drug delivery. Therefore an accurate, in-vitro BBB model may greatly aid drug delivery studies. Most current models are 2D culture systems that only roughly recapitulate the BBB. This technology uses a single platform for the study of BBB functions and has the potential for high throughput screening of the permeability of drugs and their effect on neuronal growth and function.  

Technology

This invention is a microfluidic device that consists of four channels: two for 3D hydrogels and two for culture media. The two hydrogel channels contain collagen type I and include primary rat astrocytes and neurons respectively. The hydrogel solutions are allowed to polymerize in a CO2 incubator. The fluidic channels are incubated with collagen to promote cell adhesion before being seeded with human endothelial cells. A number of studies confirm the success of this technology. Immunohistochemistry showed that astrocytes, neurons, and endothelial cells were able to grow, express cellular markers, and display the correct morphological characteristics of each of the three cell types. The endothelial cells form a monolayer barrier with accurate characteristics of the BBB, such as expression of the junction protein VE-cadherin. Neurite growth was confirmed with IMARIS software, and calcium imaging showed synaptic connectivity. Lastly, tests demonstrated size-selective permeability of the endothelial monolayer that may be altered by chemically stimulating the endothelial cells or by adding other cellular components such as pericytes. This technology offers a number of advantages, including potential for high throughput screening, low cost, the interaction of endothelial cells, astrocytes, and neurons, real-time visualization, precise control over spatiotemporal parameters, and a reduction in the amount of media, cells and chemicals required. 

Advantages

  • 3D microenvironment that is optimized for multi-cellular co-culture to more effectively model the cellular organization that is crucial for cellular processes in-vivo
  • Microfluidic system allows high throughput screening, lower costs, reduced volume of reagents and biological samples, and control of spatiotemporal parameters 
  • Real time visualization 
  • Permeability may be altered