Cell monolayer integrity was examined using an electric cell-substrate impedance sensing (ECIS) system (Applied Biophysics, Troy, NY, USA) as described (7 (link),28 (link)–30 (link)). HUVECs (20,000 cells/well) were inoculated into 96W10idfECIS array plates (Applied Biophysics) pre-coated with 10 nM cysteine and 0.1% gelatin, and cultured in ECM. The resistance of each electrode was monitored at 4000 Hz. After culturing for 48 h, the resistance reached a plateau, indicating 100% confluence. Cells were treated with 2.5, 5, or 10 μM of Kyn or ILA. Additional cells were treated with the vehicle control (DMSO, 0.02% v/v). Changes in resistance were monitored for additional 25 h.
Protocol detail
Monitoring Cell Monolayer Integrity with ECIS
Partial Protocol Preview
This section provides a glimpse into the protocol.
The remaining content is hidden due to licensing restrictions, but the full text is available at the following link:
Access Free Full Text.
Corresponding Organization :
Other organizations : Meriter Hospital, University of Wisconsin–Madison, UnityPoint Health Meriter, Shanghai First Maternity and Infant Hospital, Guangdong Medical College
Top 5 similar protocols
Variable analysis
independent variables
- Kyn concentration (2.5 μM, 5 μM, 10 μM)
- ILA concentration (2.5 μM, 5 μM, 10 μM)
- Changes in resistance of cell monolayer over time
- Cell type (HUVECs)
- Seeding density (20,000 cells/well)
- Culture medium (ECM)
- Pre-coating of cell culture plates (10 nM cysteine and 0.1% gelatin)
- Measurement frequency (4000 Hz)
- Negative control: Vehicle control (DMSO, 0.02% v/v)
Annotations
Based on most similar protocols
Etiam vel ipsum. Morbi facilisis vestibulum nisl. Praesent cursus laoreet felis. Integer adipiscing pretium orci. Nulla facilisi. Quisque posuere bibendum purus. Nulla quam mauris, cursus eget, convallis ac, molestie non, enim. Aliquam congue. Quisque sagittis nonummy sapien. Proin molestie sem vitae urna. Maecenas lorem.
Sign up for free or login to display all annotations
As authors may omit details in methods from publication, our AI will look for missing critical information across the 5 most similar protocols.
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!