EMD: Calbiochem - Product Spotlight/InnoCyte Laminin-Based 96-well Cell Invasion Assay

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InnoCyte™ Laminin-Based 96-Well Cell Invasion Assay

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Assay Features | Related Products

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The InnoCyte™ Laminin-Based 96-Well Cell Invasion Assay (Cat. No. CBA043) is designed to determine the invasive capacity of cells employing laminin as an invasion barrier. This kit can also be used to screen potential anti-metastatic agents in vitro in a convenient, high-throughput, 96-well format. The 8 mm membrane pore size is appropriate for a variety of adherent cells including epithelial, endothelial, and fibroblast cells.

Invasion is the process whereby tumor cells migrate into the surrounding stroma and disseminate via lymphatic or vascular channels to distant organs. Metastasis is the spread of tumor cells from the primary site to form secondary tumors at other sites in the body. To initiate the metastatic process, neoplastic cells must first penetrate the basement membrane and then invade the interstitial stroma by active proteolysis. Tumor invasion and metastasis are the major causes of treatment failure and death in many cancer patients.

Laminins are structural components of the basement membrane and play a key role in cell adhesion, migration, differentiation and proliferation of several cell types. They also act as barriers to the passage of cells (with the exceptions of macrophages and lymphocytes) and form a continuous sheet around epithelial structures. Laminin itself can serve as an effective barrier for non-invasive cells and is employed in various types of cell-based assays. The basement membrane is compromised by tumor cells during invasion and metastasis when the laminin layer is degraded and penetrated.

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Invasive capacity of various cell lines. HT-1080 (human fibrosarcoma), SK-MEL028 (human melanoma), HeLa (human adenocarcinoma), MCF-7 (human breast adenocarcinoma), and NIH-3T3 (murine fibroblast) cells were assessed for invasiveness using the InnoCyte™ Laminin-Based 96-Well Cell Invasion Assay. Cells were allowed to migrate/invade the laminin layer in the presence of 10% serum for 18 h. The results indicate that HT-1080 and SK-MEL028 cells exhibits strong invasiveness, while HeLa, MCF-7, and NIH3T3 cells do not significantly invade the laminin layer.

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Assay Features:

•	96-well cell culture insert contains a polycarbonate membrane coated with a laminin protein layer
•	Easily determine the invasive capacity of cells or cell lines
•	Invasive cells that degrade the laminin layer pass through the membrane and are detected with Calcein-AM
•	Convenient detection; no scraping necessary
•	The relative number of invasive cells are assessed using a fluorescence plate reader

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