Human aortic endothelial cells (HAECs) were obtained commercially (GIBCO, C0065C or PromoCell, C-12271) and cultured according to the manufacturer’s standard protocol. HAECs were seeded at various density to meet the purposes of different studies, and maintained in EC medium containing growth supplements (EBM-2, Lonza, CC-3516, CC-4176 or PromoCell, C-22011) with 2% FBS. HAECs between passage 6 and 7 were used for experiments. For shear experiments, confluent HAECs in 100 mm dishes were exposed to steady laminar shear (LS, 15 dyn/cm2), or oscillatory shear (OS, ±5 dyn/cm2) conditions for 2 days using a cone-and-plate shear device as previously described [20 (link), 21 (link)]. For shear experiments with gain of function or loss of function components, the commercially available Ibidi pump system (Ibidi, Germany) was used to reduce reagent use, and set up according to manufacturer instructions. LS (15 dyn/cm2) or OS (±5 dyn/cm2) was applied for 2 days to 80% confluent HAECs cultured overnight on microchannel slides (µ-Slide I0.4 Luer, Ibidi, Germany) coated with 40 μg/ml collagen (Collagen Type I, BD Biosciences 35–4236).
Slide 1 0.4 luer
Manufactured by Ibidi
Sourced in Germany, United States
The µ-Slide I 0.4 Luer is a laboratory slide designed for microscopic observation. It features a channel with a height of 400 micrometers and a luer connector for fluid connection. The product is suitable for various cell culture and microscopy applications.
Automatically generated - may contain errors
3 protocols using slide 1 0.4 luer
1
Shear Stress Conditioning of Aortic Endothelial Cells
2
Endothelial Cell Interaction Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
ES-HMVECs were grown to confluence in a flow chamber (µ-Slide I 0.4 Luer; Ibidi, Madison, WI, USA) coated with collagen and fibronectin as described previously [49 (link)]. After incubation with 1000 ng/mL doxycycline for 5 h, a pool of human PBMCs (5 × 105 /mL) was infused into a flow chamber at one dyn/cm2 shear stress for 5 min.
3
HUVEC Response to Low Fluid Shear Stress
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
An Ibidi pump system (Ibidi, Munich, Germany) was used to provide low-fluid shear stress (L-FSS). HUVECs were seeded onto a µ-Slide I 0.4 Luer (Special channel slides of Ibidi) and incubated for 24 h to form a monolayer. Laminar shear stress was created by attaching channel slides to a peristaltic pump. The experiment was conducted under the following conditions: 6-mbar pressure, 2.5 ml min−1 flow rate, and a shear stress of 4 dyn/cm2 HUVECs in the control group were exposed to shear stress for 24 h. The drug intervention group was cultured with RRP [(Rg1 100 μg + R1 100 μg + PCAD 30 μg) ·mL−1] for 1 h before being exposed to shear stress.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!