Transwell chemotaxis assay

Migration of cells in response to stimuli is responsible for several physiological functions mostly in the case of inflammatory responses and immune functions. The transwell chemotaxis assay is useful to study mechanisms of migration during chemotaxis. The main purpose of this assay is to determine if a molecule of interest exhibits chemotactic activity. Molecules that... 

FIGURE 3 Transwell chemotaxis assay analogous to the tumor microenvironment. Chemotaxis of tumor and endothelial cells expressing CXCR4 in response to CXCL12. The presence of the tumor outcompetes endothelial cells for CXCL12-mediated cellular migration. 

Prepare the chemotaxis plate by adding 600 pL of appropriate chemokine dilution to the bottom wells which either do not contain transwell inserts or have had the transwell inserts removed to another 24-well plate. Each point is performed in duplicate and two wells should be designated as negative controls and receive 600 pL of chemotaxis media only. These wells are used to determine the assay background. The plate may be placed at 37°C, 5% C02 to maintain temperature while preparing cells. 

The ultimate cellular response following chemokine receptor activation is chemotactic migration, i.e., chemotaxis of cells (Moser, Wolf, Walz, Loetscher, 2004). Traditionally, this is measured with an in vitro transwell assay, where suspension cells (leukocytes) are placed in droplets on top of a filter, which on its lower side is in contact with medium containing... 

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