In parallel with the Oris™ assay plate, the Reference point near the scratch in each well, and phase images were captured ofĮach scratch to document the pre-migration area of the cell-free Detection Zone. Using a 1000µL pipette tip, and serum-containing media was added to each well. Once the confluent monolayer was formed,Ĭells were serum-starved for 24 hours, then cell monolayers were scratched Following incubation, wells were rinsed, and MDA-MB-231Ĭells (500,000 cells/2mL) were seeded into each well of the Costar® plate. (Trevigen) and incubated overnight at 37 oC/5% CO 2. The Oris™ assay plate was then placedĪt 37 oC/5% CO 2 to permit cell migration.Įach well of a Costar® 6-well plate was coated with 9µg/mL collagen I Were captured, using a Zeiss Axiovert microscope with an attached CCD camera, immediatelyįollowing stopper removal to document the pre-migration area of the cell-free Detection Were serum-starved for 24 hours, stoppers were removed, and media was replaced
Once the confluent monolayer was formed, cells MDA-MB-231 human breast epithelial cells (25,000Ĭells/100µL) were seeded into all test wells of the Oris™ assay plate. Following incubation, wells were rinsed and Oris™Ĭell Seeding Stoppers removed stoppers were inserted. Oris™ Cell Migration Assay: Each well of a 96-well Oris™ TC plate wasĬoated with 9µg/mL collagen I (Trevigen) and incubated overnight at 37 oC/5%CO 2. Schematic of Oris Cell Migration AssayĬMA and the scratch assay were performed in parallel on the same day. Furthermore, ECM integrity is assessed inįigure 1. Oris™ CMA and the scratch assay in assessing cell migration. This application note offers a direct comparison of the Result of uniformly sized Detection Zones, and the underlying ECM is not Reproducibility is greater in the Oris™ CMA compared to the scratch assay as a The center of each well, into which cells are permitted to migrate. To adhere, the silicone stoppers are removed to reveal an unseeded region in Silicone stoppers that exclude cells from the central Detection Zone of the
The Oris™ CMA (Figure 1) uses a 96-well plate populated with Oris™ Cell Migration Assay (CMA) was designed to address the limitations of the Furthermore, the process of scratch formation has been shown to damage the underlying extracellular matrix (ECM). However, methods for creating the scratch vary from lab to lab and results can be highly variable. The scratch assay is straightforward to perform and is inexpensive.
Cell migration can be assessed by comparing images captured at the onset of the scratch creation and at user-defined intervals during scratch closure. The scratch assay is performed by creating a cell-free gap, or “scratch”, on a confluent cell monolayer upon which cells at the edge of the opening move inward to close the scratch. One assay commonly used to study cell migration in vitro is the scratch assay. In addition, cell migration is involved in tumor metastasis and atherosclerosis. Cell migration is integral to many physiological processes, including embryonic development, tissue regeneration, and wound healing.
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