Cell migration is a highly integrated, multi-step process. Dysregulated cell migration has been implicated in cancer, macular degeneration and diabetic wound healing. Understanding the processes involved in cell movement can provide important insight into the management of multiple disease states. Traditional methods to monitor 2-D closure have utilized scratch assays but these can produce inconsistent detection zones and are not readily compatible with high-throughput screening strategies. The Oris™ Cell Migration Assay utilizes unique cell-seeding stoppers to create a detection zone for visualizing cell migration/cell motility and invasion. The Oris assay is well suited for testing compounds that inhibit undirected tumor cell migration. This 96-well format assay can be used for screening with medium throughput rates with high accuracy.
Utilizing the human medulloblastoma cell line, Daoy, FGF activation has been shown to drive the activation of PI3K/AKT, STAT, p38 MAPK, JNK, SRC and RSK2 pathways which collectively play multiple roles in cell survival, growth, migration, differentiation, and metabolism.
Daoy cells are seeded onto collagen I-coated multi-well plates equipped with a barrier that prevents cell adherence to the center of each well. After 24 hours, the barrier is removed revealing a clear region in the center of the well. Cells are prestained with Hoechst prior to the assay. bFGF stimulation followed by the addition of test compounds are imaged at 0 and 24hrs, during which the Daoy cells migrate into the detection zone. Highlights:
- 96-well format
- 8-point concentration range to generate an EC50 or as single concentration for library screening
- High Content Imaging readout
- Can also be developed using alternative tumor cells
