SynVivo’s  SynBBB 3D blood brain barrier model recreates the in vivo microenvironment by emulating a histological slice of brain tissue cells in communication with endothelial cells across the blood brain barrier (BBB). Shear-induced endothelial cell tight junctions, which cannot be achieved in the Transwell® model, are easily achieved in the SynBBB model using physiological fluid flow. Formation of tight changes can be measured using biochemical or electrical analysis (assessing changes in electrical resistance) with the SynVivo Cell Impedance Analyzer. Interactions between brain tissue cells and endothelial cells are readily visualized in the SynBBB assay. Transwell models do not allow real-time visualization of these cellular interactions, which are critical for understanding of the BBB microenvironment.

SynBBB is the only in vitro BBB model that allows:

  • Accurate in vivo hemodynamic shear stress
  • Real-time visualization of cellular and barrier functionality
  • Significant reduction in cost and time
  • Robust and easy to use protocols

Schematic of the BBB Model. Apical chamber (outer channels) are for culture of vascular (endothelial cells) while basolateral chamber (central chamber) are for culture of brain tissue cells (astrocytes, pericytes, neurons). Porous architecture enables communication between the vascular and tissue cells.

The SynBBB system is a highly versatile platform for investigation of:

  • Tight junction proteins: Determine the levels of tight junction proteins namely zonula occludens, claudins and occludins which regulate the BBB.
  • Transporter proteins: Analyze functionality of transporter proteins (e.g. Pgp) in normal and dysfunctional BBB.
  • Drug permeability: Evaluate real-time permeability of therapeutics and small molecules across the endothelium of the BBB.
  • Inflammation: Understand the underlying mechanisms of  inflammatory responses on the regulation of the BBB.
  • Cell migration: Visualize and quantify in real-time migration of immune cells across the BBB.
  • Omic changes: Perform genomic, proteomic and metabolic analysis on normal and dysfunctional BBB.
  • Neurotoxicity: Analyze toxicity effects of chemical, biological and physical agents on the cells of the BBB.
  • Neuro-oncology: Investigate effects of the tumor cells on the BBB.

Depending on your research needs you can select from the “basic” SynBBB model or a “TEER-compatible” SynBBB configuration.

Basic SynBBB Configuration

Analyze experiments using real-time imaging, biochemical and molecular biology methodologies.

TEER Compatibile SynBBB Configuration with the SynVivo Cell Impedance Analyzer

Analyze experiments using real-time trans-endothelial electrical resistance (TEER) including real-time imaging, biochemical and molecular biology methodologies.

SynBBB 3D Model Kit Components

All the basic components required to run the SynBBB assay can be purchased in a kit format. Depending on individual research needs you can select from the “basic” or “TEER-compatible” configurations of the SynBBB chip. All accessories including tubing, clamps, needles and syringes are included. Starter kits will also include the pneumatic priming device (required for running SynBBB assays) and the cell impedance analyzer (required for collecting SynBBB TEER measurements).

Qty Description Cat#
10 SynBBB Chips (basic or TEER capable configuration) 102005 or 102015
100ft Tygon Tubing (0.02″ ID x 0.06″ OD) 201005
25 Slide Clamps 202003
50 Blunt Tip Needles (24ga x 0.5″ long) 204002
50 1mL Syringes with Luer-Lock Tips 203004
1 SynVivo Pneumatic Primer Device (in starter kits only) 205001
1 SynVivo Cell Impedance Analyzer (in TEER starter kit only) 304001

 

SynVivo used to create the first neonatal BBB model on a chip

Researchers at Temple University used the SynVivo® SynBBBTM cell-based in vitro assay platform to model the attributes and functions of the neonatal stage blood-brain barrier (BBB) [1]. The SynBBB model closely mimics the in vivo microenvironment including three-dimensional morphology, cellular interactions and flow characteristics on a microfluidic chip. This work marks the first dynamic in vitro neonatal BBB model that offers real time visualization and analysis and is suitable for studies of BBB function as well as screening of novel therapeutics.

“The work is important because studies of neonatal neuropathologies and development of appropriate therapeutics are hampered by a lack of relevant in vitro models of the neonatal blood-brain barrier,” said Dr. Sudhir Deosarkar, the lead author of this paper.

In the SynBBB assay, which includes a tissue compartment and vascular channels placed side-by-side and separated by an engineered porous barrier, the researchers were able to co-culture neonatal rat brain endothelial cells and rat astrocytes under physiological conditions observed in vivo. The endothelial cells formed a full lumen and exhibited tight junction formation which increased under co-culture with astrocytes. The permeability of small molecules in the developed model was found to in excellent agreement with in vivo observations.

“The real-time visualization capabilities of the SynBBB co-culture platform allowed, for the first time, visualization of astrocyte end-feet and endothelial cell interactions in anin vitro model,” said Prof. Mohammad Kiani who is the senior author of the paper. “This is a unique capability and will help us to understand and develop therapeutics for several developmental disorders and diseases of the brain.”

astrocyte-communications

The PLOS ONE paper shows that in contrast to transwell models, the SynBBB model exhibits significantly improved barrier characteristics similar to in vivo observations.

1A Novel Dynamic Neonatal Blood-Brain Barrier on a Chip. S. Deosarkar, B. Prabhakarpandian, B. Wang, J.B. Sheffield, B. Krynska, M. Kiani. PLOS ONE, 2015, DOI: 10.1371/journal.pone.0142725

The SynBBB 3D model has been validated in various BBB Assays

Mono-Culture Assays

Shear-induced endothelial cell tight junctions, which cannot be achieved in the Transwell® model, are easily achieved in the SynBBB assay using fluid perfusion. Formation of tight changes can be measured using biochemical or electrical analysis (assessing changes in electrical resistance) with the SynVivo Cell Impedance Analyzer.

Primary endothelial cells are cultured in the vascular channel under physiological fluid flow. Cells are stained for tight junction markers highlighting the increase under fluid flow compared to static conditions. The Cell Impedance Analyzer system is used to measure increases in Ohmic resistance (TEER), associated with the formation of tight junctions.

Top Left Panel: Phase Contrast imaging of brain endothelial cells cultured in the SynBBB model. Bottom Left Panel: Calcein AM and Ethidium homodimer-1 labeled brain endothelial cells indicating a highly viable population of cells in the SynBBB model. Right Panel: Plot highlighting the importance of flow on brain endothelial cells with increased TEER.

Co-Culture with Tissue Cells

Interactions between brain tissue cells and endothelial cells are readily visualized in the SynBBB assay. Transwell models do not allow real-time visualization of these cellular interactions, which are critical for understanding of the physiological environment.

Endothelial cells are cultured under flow in the vascular channel, and the tissue chamber is cultured with primary brain cells, such as astrocytes. Increases in Ohmic resistance across the barrier, measured with the Cell Impedance Analyzer, are associated with tight junction formation across the BBB. Endothelial cells co-cultured with astrocytes form significantly tighter cell junctions compared to mono-cultured endothelial cells.

Left Panel: CD-31 (green) stained endothelial cells and GFAP (red) stained astrocytes. All nucleus are stained with DAPI (blue). Right Panel: Plot highlighting increased TEER with co-culture of endothelial cells and astrocytes.

Real-Time Permeability Assays

Unlike BBB models which are arranged in top to bottom architecture (i.e., Transwell), small molecule transport can be assessed and quantified in real-time across the SynBBB system due to its side-by-side architecture.

A fluorescently-labeled drug molecule of interest is perfused through the vascular channels at physiological  flow rate. Real-time videos are acquired and analyzed to calculate the rate of permeability into the tissue chamber. Different rates of permeability is observed across the BBB due to tight junctions of endothelial cells.

Time-lapse imaging of permeability of small molecules across a tightly formed BBB.

Time-lapse imaging of permeability of small molecules across a leaky BBB.

Real-Time Tight Junction Modulation

SynBBB can be used to model inflammation responses. A pro-inflammatory compound, such as TNF-α, is added to mono-cultured endothelial cells to modulate the tight junctions, followed by a period of recovery under perfusion flow. Electrical resistance measurements provide a non-invasive method for real-time monitoring of tight junctions.

Modulation of Inflammation responses in SynBBB model. TNF-alpha induced leakiness in the BBB measured by changes in the resistance across the endothelial cells. Removal of TNF-alpha followed by media perfusion under physiological flow conditions enables recovery of the tight junction leading to increased tight junction formation. Static cells maintain a constant resistance due to lack of tight junctions.

3D TISSUE MODELS AND SERVICES-BROCHURE

SynBBB 3D Model (basic configuration) Technical Manual
SynBBB 3D Model (TEER configuration) Technical Manual

SynBBB 3D Model – Starter Kits

Includes consumables (10 chips, 100ft tubing, 25 slide clamps, 50 blunt tip needles and 50 1 ml syringes). Starter kits will also include the pneumatic priming device (required for seeding cells).

Cat#
Product Name
Literature
Price
402002
SynBBB 3D Model Starter Kit (basic configuration)

$1,260 Add to cart

Note: Does not include other required consumables such as cells, media and matrix.
Laboratory equipment required includes incubators, inverted microscopes, and syringe pumps.

SynBBB 3D Model – Assay Kits

Includes consumables (10 chips, 100ft tubing, 25 slide clamps, 50 blunt tip needles and 50 1 ml syringes).

Cat#
Product Name
Literature
Price
402001
SynBBB 3D Model Assay Kit (basic configuration)

Note: Does not include other required consumables such as cells, media and matrix.
Laboratory equipment required includes incubators, inverted microscopes, and syringe pumps.

SynBBB 3D Model – Chips

Purchase single chips.

Cat#
Product Name
Literature
Price
102005-SB
SynBBB 3D Model Chip (basic configuration)
102015-SB
SynBBB 3D Model Chip (TEER configuration)

Note: Does not include other required consumables such as tubing, clamps, syringes, needles, cells, media and matrix.
Laboratory equipment required includes incubators, inverted microscopes, and syringe pumps.