General considerations for optimizing cell migration and invasion assays incorporating ECM

The extracellular matrix (ECM) is a complex network of proteins and polysaccharides that provides structural and biochemical support to surrounding cells. It includes components like collagen, elastin, fibronectin, and laminin, which form a scaffold that influences cell behavior, including adhesion, migration, and differentiation[1]. In cancer cell invasion and metastasis assays, the ECM mimics the in vivo environment, making it crucial for accurate and reproducible results.

For optimizing these assays, consider the following technical guidance:

1. Matrix composition:

   Choose the appropriate ECM components based on the specific cancer cell type and the biological question. For instance, Matrigel, a basement membrane matrix, is commonly used for invasion assays due to its rich composition of ECM proteins [2].

2. Coating consistency:

   Ensure uniform coating of the ECM on assay plates. This can be achieved using Agilent BioTek automated liquid handling systems, which provide precise and reproducible dispensing of ECM solutions [2].

3. Cell seeding density:

   Optimize cell seeding density to prevent overcrowding, which can lead to nutrient depletion and altered cell behavior. Characterization of single-cell migration and invasion requires a low cell density to track and measure individual cells, whereas collective cell migration studies typically rely on a uniform confluent layer of cells [2].

By carefully considering these factors, you can enhance the reliability and accuracy of your cancer cell invasion and metastasis assays.