The Oris™ Cell Invasion Assays offer a more versatile method for quantitating and imaging cells invading through an extracellular matrix (ECM) compared to a Boyden chamber device or membrane cell culture insert. Using Oris™ Cell Seeding Stoppers to create an invasion zone at the bottom of a microplate well, researchers can detect cell invasion in a 3-D Collagen I or Basement Membrane Extract (BME) environment and monitor cell movement in real-time (Fig. 1).
Features & Benefits of the Oris™ Cell Invasion Assays
Cell invasion assays have become a critical technique in screening compounds that can inhibit metastasis or promote angiogenesis1. True cell invasion is indicated by the ability of an adherent cell line such as HT-1080 human fibrosarcoma cells to use proteases and digest through an ECM component which can mimic an in vivo environment2,3. The Oris™ Cell Invasion & Detection Assay allows researchers to study the specific mechanisms involved with cell invasion (Fig. 3 & 4) by making it easy to stain proteins associated with proteolysis and cytoskeletal structures in a single well. This cellular activity can be quantitated using a plate reader (Fig. 2) as well as imaged with a microscope or digital imaging system (Fig. 1).
Figure 1. HT-1080 Cell Invasion Microscopic Data
Figure 2. HT-1080 Cell Invasion Plate Reader Data
Figures 1 & 2: Invasion of HT-1080 cells into BME: HT-1080 cells, serum starved for 18 hours, were seeded (50,000 cells/well) on an ECM coated plate (4 mg/ml). Stoppers were removed and BME (+/- 10% FBS) was overlayed on the cells. Following a 48 hour incubation the cells were labeled with Calcein AM and imaged by using a Zeiss Axiovert microscope (5X magnification). Fluorescence in the analytic zone was quantified by using a plate reader. Each column represents the mean +/- SEM of at least 22 wells. A non-invasive cell line, 3T3-Swiss albino, served as the negative control.
Figure 3. Anti-Cortactin Staining Data
Figure 4. Anti-Cathepsin B Staining Data
Figures 3 &4: Anti-cortactin and anti-cathepsin staining of HT-1080 cells: F-actin is present in all eukaryotic cells, it is a major component of the cytoskeleton and functions to define and maintain the cell shape. Cortactin is a cellular protein that is present in and promotes the formation of lamellipodia and invadopodia. These structures propel cells over surfaces as they move toward a target. Results depicted above demonstrate distinct areas of Cortactin and F-actin colocalization characteristic of invadopodiaRef 2,4. Cathepsin B is a lysosomal protease that facilitates cell invasion by degrading the surrounding extracellular matrix. Results depicted above demonstrate distinct areas of lysosomal and peripheral cellular expression of Cathepsin B characteristic of protease expression observed in cancer cell invasionRef 5.
ORDER the Oris™ Cell Invasion Assay Kits
| Product Number | Product Description | Package Size | Price |
|---|---|---|---|
| CIA101CC | Oris™ Collagen I Cell Invasion Assay Kit, 1-pack: Oris™ Collagen I coated, 96-well plate with Oris™ Cell Seeding Stoppers, 1 Oris™ Detection Mask, 1 Oris™ Stopper Tool, 1 Oris™ Collagen I Stock Reagent, 4 mL Oris™ 10X PBS Buffer, 1 mL |
1-pack | $350 |
| CIA200CC | Oris™ Collagen I Cell Invasion Assay Kit, 2-pack: Oris™ Collagen I coated, 96-well plate with Oris™ Cell Seeding Stoppers, 2 Oris™ Detection Mask, 1 Oris™ Stopper Tool, 1 Oris™ Collagen I Stock Reagent, 2x 4 mL Oris™ 10X PBS Buffer, 2x 1 mL |
2-pack | $650 |
| CIA101DE | Oris™ Cell Invasion Assay Kit, 1-pack: Oris™ Compatible, 96-well plate, 1 Oris™ Cell Seeding Stoppers, 96 Oris™ Detection Mask, 1 Oris™ Stopper Tool, 1 Oris™ Basement Membrane Extract (BME), 1 x 5 mL |
1-pack | $480 |
| CIA200DE | Oris™ Cell Invasion Assay Kit, 2-pack: Oris™ Compatible, 96-well plates, 2 Oris™ Cell Seeding Stoppers, 2 x 96 Oris™ Detection Mask, 2 Oris™ Stopper Tool, 2 Oris™ Basement Membrane Extract (BME), 2 x 5 mL |
2-pack | $870 |
| CMA.IRT | Oris™ Stopper Tool |
1 | $20 |
| CMA.MM | Oris™ Detection Mask |
5 | $35 |
* Patent Pending
Oris™ is a trademark of Platypus Technologies, LLC
Transwell® is a registered trademark of Corning, Inc.
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References:
1. Stetler-Stevenson, W.G. et al., Tumor Cell Interactions with Extracellular Matrix During Invasion and Metastasis. (1993) Annu. Rev. Cell. Bio. 9:541-73.
2. Waever, A.M. Invadopodia: specialized cell structures for cancer invasion (2006) Clin. Exp. Metastasis 23:97-105.
3. Friedl, P. and Bröcker, E. B. The biology of cell locomotion within a three-dimensional extracellular matrix (2000) Cell. Mol. Life Sci. 57: 41-64.
4. Furmaniak-Kazmierczak et al; Formation of extracellular matrix-digesting invadopodia by primary aortic smooth muscle cells (2007) Circulation Research 100:1328-1336.
5. Hulkower, KI et al; Fluorescent microplate assay for cancer cell-associated cathepsin B (2000) Eur. J. Biochem 267:4165-4170.
