The mobility of leukocytes plays an important role in immune response to maintain the homeostasis of microenvironment in vivo. Meanwhile, the leukocyte migration is related to infection and inflammation, which could cause serious complications and interfere to the clinical treatment of many diseases. Thus, the study of leukocyte migration is significant and essential. Among all methods used to study cell migration of leukocyte, the leukocyte migration agarose test (LMAT) is a convenient and simple detection method. This technique also has other advantages such as cheap and easy to set up ect. The LMAT assay can be used to mimic that monocytes and macrophages can increase the migration of neutrophils in the presence of glucocorticoid in vivo. Other factors such as CXXL8, which could influence the migration of neutrophils, also can be studied by this technique.
The leukocyte migration agarose test (LMAT) is relatively simple and easy, compared to other methods used to research cell migration. A pre-cast thin agarose layer on a tissue culture dish is prepared and circular wells into the agarose layer are cut out in defined distances. Leukocytes are loaded into one of these wells, whereas the adjacent holes are filled with different media that contain different kinds of factors such as chemotactic factors and inhibitors. Then, the leukocytes will migrate below the agarose layer toward the different media reservoirs and the distance of migration toward the test substance is detected and scored by living image system. The migratory distance that leukocytes toward the test substance can be divided by the migratory distance toward the control medium to define the migratory index. Compared the distance and cell numbers of leukocytes that migrate to different media, we can assess the chemotaxis induced by different factors accurately.
Figure 1. The schematic diagram of leukocyte migration agarose test (LMAT) assay
The leukocyte migration agarose test can be influenced by many experimental conditions such as cell number of leukocytes that loaded into the well, the distance between wells, the incubation time, contamination and so on. With more than ten years of experience, Creative Proteomics take all these factors into consideration to ensure the sensitivity, accuracy, consistence and reproducibility of the results. Experimental protocol of LAMT can also be optimized according to your requirement and purpose of the research.
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