and inflammation response
Transmigration of leukocytes from the lumen of the blood vessels to the underlying tissue is a key process in immunosurveillance and in the response to inflammation and injury in the normal function of the immune system. However, in inflammatory diseases and cancer, transmigration of cells to underlying tissues is a gateway to metastasis and morbidity. The significance of transmigration of unwanted cells to underlying tissues and through the blood brain barrier from the vasculature in disease states makes it an important target for drug development. As such, transmigration is a prime target for inflammation and oncology drug discovery.
The BioFlux system can be used to study live cell transmigration in a physiologically-relevant assay using primary endothelial cells and human peripheral blood mononuclear cells. Studying transmigration under shear flow is necessary to maintain physiologically-relevant conditions for this biological process in blood vessels. The system can be used to study the dynamic process of transmigration using time-lapse microscopy under flow and post-experiment image analysis to determine migration distance. The system can also be used to examine 24 different monolayers at one time, which is useful for determining IC50 concentrations under shear flow or screening multiple compounds under the same experimental conditions.
Under shear stress with a BioFlux 1000z, human umbilical vein endothelial cells (HUVECS) and human monocytic leukemia cell line THP-1 were treated with anti-inflammatory acetylharpagide to model wound healing. Adherent cells (A) and transmigrated cells (B) are marked with white arrowheads. Acetylhapagide significantly inhibited wound healing (C) (You et al., 2014).
Regulation of chemokine-dependent adhesion of dendritic cells (DCs) with lymphatic endothelial heparan sulfate was analyzed by using a BioFlux. Lymphatic endothelial cells (LECs) were seeded into chamber and treated with antibodies or siRNA. DCs were labeled and seeded into the chamber. Channels were imaged after nonadherent cells were washed away (Yin et al., 2016).
Key BioFlux Advantages For Transmigration Studies
Study live cell transmigration in a physiologically-relevant assay using primary endothelial cells, human peripheral blood mononuclear cells, and more
Utilize time-lapse microscopy under flow and post-experiment image analysis to determine migration distance
Examine dozens of different monolayers at one time in order to determine IC50 concentrations or screen multiple compounds under the same experimental conditions
With the BioFlux heated stage insert, alter the temperature of experiments to analyze cells at ambient temperature or 37 C (a temperature permissive for transmigration)