Vena8 Endothelial+ Biochip: for cell-primary endothelial cell studies and mimicking physiological flow
Vena8 Endothelial+ biochips contain 8 parallel enclosed microcapillaries for culturing primary endothelial cells and continuous flow cell-based assays. Primary endothelial cells are cultured, and cell suspensions may then be injected using Cellix’s microfluidic pumps which supports a range of shear stresses / shear flow rates for dynamic flow-based assays. Vena8 Endothelial+ biochips are supplied in packs of 10, facilitating 80 experiments per pack.
Vena8 Endothelial+ Features
20x, 40x, 60x, 100x short working distance magnification microscopy; 60x, 100x oil- immersion microscopy.
Wider channels for easy seeding of cells.
Primary endothelial cell monolayer obtained in 3 hours.
Compatible with brightfield / phase contrast / fluorescent / confocal microscopy.
Suitable for culturing wide range of primary endothelial cells.
Suitable for whole blood and blood cells analysis (e.g. leukocytes, platelets)
Biochip plastic is optically clear permitting detailed microscopy studies.
0.05–200 dyne/cm2 shear stresses / shear flow rates easily obtained and controlled
by the Mirus Evo nanopump, ExiGo, UniGo and 4U microfluidic pumps.
Shear stress / shear flow rate may be pre-set to be incrementally increased during an
Real time imaging under flow conditions.
Cell culture of endothelial cell monolayers for shear flow experiments.
Cell-cell adhesion shear flow assays on endothelial cell monolayers for studies such as:
Cardiovascular / Atherosclerosis: primary monocyte adhesion to coronary artery endothelial cells.
Inflammation: THP-1 monocyte adhesion to HUVECs under flow.
Asthma / COPD: eosinophil adhesion studies to human lung microvascular endothelial cells (HMVEC-L).
Oncology - Breast Cancer: MDA cells adhering to TNF-alpha stimulated HUVECs under shear flow.
Performance and Technical Specifications
*Considering human whole blood with a viscosity of 4.5 cP.
**Given for the flow of distilled water in a microcapillary with dimensions: 400 μm (W) x 100 μm (D) x 28 mm (L).
Vena8 Endothelial+ Biochip, Protocol #1: coating and cell seeding in Vena8 Endothelial+ biochips
Cellix Vena8 Endothelial+ biochip is coated using a standard yellow tip pipette, by dispensing approximately 12 μL of protein (e.g. fibronectin, 100 μg/mL) into each microchannel. Note the excess of liquid on the entrance and exit ports.
The Vena8 Endothelial+ biochip is then placed in a humidified box which remains open for 1–1.5 hours in the CO2 incubator. Alternatively, the biochip may be placed at 4°C for overnight coating.
After the incubation period, add 5 μL of 1.5 x 106 /100 μL (≅ 15 x 106/mL)of endothelial cells gently into each channel.
Note: concentration specified is for primary HUVEC.
The biochip is kept in the CO2 incubator for 15–20 minutes for the cells to adhere. Observe the biochip under a microscope and top up all the reservoirs with 50 μL of media. Keep the biochip for 1.5–2 hrs in the CO2 incubator.
Vena8 Endothelial+ Biochip Protocol #2: executing cell rolling, adhesion and migration assays under shear flow with Vena8 Endothelial+ biochips (manual version — not with VenaFlux platform)
Suspension cells (e.g. T cells, monocytes, platelets) are re-suspended in culture medium at an appropriate concentration (typically 2–5 x 106/mL) in an Eppendorf tube.
Using the Cellix Mirus Evo nanopump or the ExiGo pump, 10 μL of media is dispensed from pump output cable. Following this, the output cable is inserted into a specified channel on the Vena8 Endothelial+ biochip.
Then using the Cellix Mirus Evo nanopump or the ExiGo pump, 40 μL of the media is injected through the channel at a shear stress of 40 dynes/cm2. This is done to wash the channel of cell debris. The waste is aspirated from the microwell of Vena8 Endothelial+ biochip with a pipette.
Cell sample is placed into the microwell of this channel on the Vena8 Endothelial+ biochip.
Cells are introduced into the channel, by specifying the desired shear stress on the FlowAssay software. The flow rate will be automatically calculated.
At each shear stress value, it is recommended that images of 3–5 fields of view of cell rolling and adhesion are acquired along the length of the channel.
So now you know the features and benefits of our Vena8 Endothelial+ and how they works. But, if you still have questions or want to learn more, check out our website, at wearecellix.com, or contact us at firstname.lastname@example.org.