The SynRAM™ 3D Inflammation Model from SynVivo has been developed to study the entire inflammation pathway in a realistic and dynamic environment. By recreating a histological slice of co-cultured tissue and/or tumor cells with a lumen of endothelial cells, the SynVivo platform delivers a physiologically realistic model including flow and shear in a platform and enables real-time tracking of rolling, adhesion and migration processes. This model has been successfully validated against in vivo studies showing excellent correlation with rolling velocities, adhesion patterns and migratory processes (Lamberti et al 2014, Soroush et al 2016).

The SynRAM 3D inflammation model provides a realistic testing environment including:

  • Physiological shear stress within a microvascular environment
  • In vivo like vascular morphology with fully enclosed lumen
  • Co-culture capability for cell-cell interactions
  • Quantitative real-time rolling, adhesion, and migration data from a single experiment

SynRAM enables assessment of cellular interactions comprising of rolling, adhesion and migration through multiple cellular layers in one experiment, in real-time, and represents data closely correlated with in vivo results.

SynRAM’s innovative design overcomes the current limitations inherent in flow chambers or Transwell chamber based assays.  Current flow chamber designs are oversimplified, lack the scale and geometry of the microenvironment and cannot model transmigration. Similarly, Transwell chambers do not account for fluid shear and size/topology observed in vivo, the end point measurements of migration are not reproducible and do not provide real-time visualization.

SynVivo’s proprietary chip designs range from complex in vivo derived microvascular networks (obtained from digitized images) to produce realistic cellular makeup and vascular morphology resulting in varying shear and flow conditions, to simplified idealized networks designed to reproduce the cellular makeup and constant shear and flow conditions.

Idealized Network

Microvascular Network

SynRAM 3D Model Kit Components

All the basic components required to run assays using the the SynRAM model can be purchased in a kit format. Depending on individual research needs you can select from “idealized” or “microvascular” configurations of the SynRAM chip. All accessories including tubing, clamps, needles and syringes are included. Starter kits will also include the pneumatic priming device (required to run assays using SynRAM).

Kits include the following components:

Qty Description Cat#
10 SynRAM Chips (“idealized” or “microvascular”) 102008 or 105001
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 kit only) 205001

SynRAM successfully validated against in vivo data

SynRAM microfluidic chips comprising of realistic microvascular networks were used to understand the role of classical inhibitors of individual steps of the leukocyte adhesion cascade. Experimental results matched very well with in vivo data highlighting the unique ability of the platform for real-time analysis of these dynamic events in a morphologically realistic environment (Lamberti et al 2014).

Rolling velocity

Neutrophil rolling using SynRAM microfluidic chips is similar to leukocyte rolling in vivo; Box and whisker plots summarizes the comparison of leukocytes rolling velocity measured in vivo and in SynRAM chips and shows no significant difference (p=0.758; Mann-Whitney Rank Sum Test). The “+” marked in the box indicates the mean.

synvivo_charts-11

Neutrophil adhesion in SynRAM microfluidic chips is similar to leukocyte adhesion in vivo; Distribution of the number of adhered leukocytes and neutrophils as a function of distance from the nearest bifurcation in vivo in mouse cremaster muscle model and in vitro in microfluidic chips, respectively. Both histograms are skewed to the left indicating that leukocytes and neutrophils preferentially adhere near bifurcations with the peak occurring at one vessel or channel diameter from the nearest bifurcation.

Bioinspired Microfluidic Assay for In Vitro Modeling of Leukocyte–Endothelium Interactions. G. Lamberti, B. Prabhakarpandian, C. Garson, A. Smith, K. Pant, B. Wang, and M.F. Kiani. Anal. Chem., 2014, 86 (16), pp 8344–8351 DOI:10.1021/ac5018716

Investigation of the Effect of Blocking of Specific Steps of the Inflammation Pathway using Monoclonal Antibodies

Antibody blocking of specific steps in the adhesion/migration cascade downregulates other steps of the cascade; Monoclonal antibodies against E-selectin (aE-selectin), ICAM-1(aICAM-1), and PI3K (wortmannin) significantly reduced the number of rolling, adhering, and migrating neutrophils in SynRAM microfluidic devices.

Percent activity after treating cells with the respective antibody blockers in comparison to their corresponding control values.

Elucidation of the Mechanism of Protein Kinase C delta (PKCδ) in Sepsis Related Inflammation Response

The SynRAM model was used to identify the underlying mechanism of Protein Kinase C delta (PKCδ) dependent neutrophil-endothelium interactions which has been found to play a significant role in the inflammatory response.  Under physiological fluid flow conditions and using the simultaneous real-time monitoring ability of the entire inflammation process comprising of rolling, adhesion and migration, they found PKC𝛿 was a critical regulator of human neutrophil adhesion and migration through human endothelial cells during inflammation.

PKCδ-TAT inhibitor significantly reduces migration of neutrophils from the vascular channels, across the inflammed endothelium (treated with TNF-α for 4 or 24 hour), into the tissue compartment in response to fMLP mediated signaling compared to untreated controls.

Immunohistochemical detection of myeloperoxidase (MPO) in representative lung tissue sections from 24 h post surgery.  Few MPO-positive cells in Sham surgery. Sepsis induces the infiltration of numerous MPO-positive cells throughout the lung parenchyma. PKCδ-TAT Inhibitor significantly reduces sepsis-induced, MPO-positive cell numbers in the lung indicating decreased neutrophil migration.

The SynRAM 3D Inflammation Model can be used in Various Cell Migration Assays

Chemotaxis and Migration

Cells are incubated in the vascular channel and chemoattractant is injected into the tissue chamber or the second vascular channel (if using the idealized network). Migration of immune cells across the engineered porous region into the tissue chamber is observed in real-time. The assay can be performed under static or flow conditions.

Vascular (endothelial cells) Mediated Rolling, Adhesion and Migration Assay

Endothelial cells cultured in the vascular channel are activated with biological or chemical agents to produce an inflammatory response followed by real-time tracking of rolling, adhesion and migration of immune cells across the endothelium into the tissue chamber.

Vascular and Tissue Co-culture Mediated Rolling, Adhesion and Migration Assay

Endothelial cells are cultured in vascular channel while tissue chamber houses smooth muscle cells, epithelial cells, fibroblast or organ specific cells (e.g. astrocytes, hepatocytes). In response to a cellular signaling and inflammation activation, rolling, adhesion and migration across the endothelium can be observed and quantified in real-time.

Rolling, adhesion and migration of WBCs across the inflamed vascular-tissue interface.

3D TISSUE MODELS AND SERVICES-BROCHURE

migration-assay-flow

SynRAM 3D Model (idealized network) Technical Manual
SynRAM 3D Model (microvascular network) Technical Manual

SynRAM 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 running SynRAM assays).

Cat#
Product Name
Literature
Price
401002
SynRAM 3D Model Starter Kit (idealized network)

$1,260 Add to cart

401004
SynRAM 3D Model Starter Kit (microvascular network)

$1,560 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.

SynRAM 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
401001
SynRAM 3D Model Assay Kit (idealized network)
401003
SynRAM 3D Model Assay Kit (microvascular network)

$1,290 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.

SynRAM 3D Model – Chips

Purchase single chips.

Cat#
Product Name
Literature
Price
102008-SR
SynRAM 3D Model Chip (idealized network)
105001-SR
SynRAM 3D Model Chip (microvascular network)

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.