Transmigration Assay
Transmigration Assay - Related Videos
03:39
Studying Lymphocyte Transmigration Across a Microvascular Endothelial Cell Monolayer
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2025
This video demonstrates an experimental investigation of lymphocyte transmigration across a blood-brain barrier (BBB) endothelium model. A monolayer of human brain microvascular endothelial cells (HBMECs) is generated in vitro, and peripheral blood mononuclear cells (PBMCs) containing lymphocytes are added to the monolayer. The cells are then incubated to allow lymphocyte transmigration, and the migrated cells are quantified using flow cytometry.
08:54
Isolation of Human Umbilical Vein Endothelial Cells and Their Use in the Study of Neutrophil Transmigration Under Flow Conditions
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Cited by 33 •
2012
This article first describes a procedure for isolating human endothelial cells from umbilical veins and then shows how to use these cells to examine neutrophil transmigration under flow conditions. By using a low-volume flow chamber made from a polymer with the optical characteristics of glass, live-cell fluorescent imaging of rare cell populations is also possible.
10:25
Assessment of Lymphocyte Migration in an Ex Vivo Transmigration System
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Cited by 3 •
2019
In this protocol, lymphocytes are placed in the top chamber of a transmigration system, separated from the bottom chamber by a porous membrane. Chemokine is added to the bottom chamber, which induces active migration along a chemokine gradient. After 48 h, lymphocytes are counted in both chambers to quantitate transmigration.
03:54
A Laminar Flow-Based Assay to Study the Interactions Between Leukocytes and Endothelial Cells
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2025
This video demonstrates a laminar flow-based assay to characterize the interaction of leukocytes and cellular partners such as endothelial cells. This method provides dynamic insights into adhesion, deadhesion, and transmigration of leukocytes when perfused over an intact and confluent layer of vascular cells.
09:41
Quantification of Monocyte Transmigration and Foam Cell Formation from Individuals with Chronic Inflammatory Conditions
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Cited by 17 •
2017
We describe a protocol to measure transmigration by monocytes across human endothelial monolayers and their subsequent maturation into foam cells. This provides a versatile method to assess the atherogenic properties of monocytes isolated from people with different disease conditions and to evaluate factors in blood which may enhance this propensity.
FAQs
The chemoattractant placed in the lower compartment establishes a concentration gradient across the barrier. Cells respond by moving toward the compartment with the attractant, allowing directed migration to be distinguished from movement that is not guided by a defined signal. Changing the gradient can therefore affect the apparent strength of chemotactic recruitment.
A porous membrane provides a controlled physical barrier through which cells must pass, whereas an endothelial monolayer models a cellular vascular barrier. The membrane emphasizes passage through pores, while the monolayer also permits examination of endothelial barrier behavior. Comparing these formats can help separate cell-migration effects from changes associated with endothelial permeability.
The assay can provide information about chemotaxis, endothelial permeability, and immune-cell recruitment, but these readouts are related rather than identical. Directed movement reflects responsiveness to the chemoattractant, passage through an endothelial layer reflects interaction with a biological barrier, and the number of cells crossing indicates the resulting recruitment under the tested conditions.
A typical workflow places the test cells in an upper compartment containing a porous membrane or endothelial monolayer and introduces a chemoattractant below it. After allowing migration to occur, investigators quantify the cells that have crossed the barrier. This arrangement creates a controlled comparison between cells exposed to the migration stimulus and the barrier condition being studied.
The number of cells recovered beyond the barrier provides a measurement of migration under the selected conditions. Interpreted alongside the type of barrier and the presence of a chemoattractant, this result can indicate differences in chemotactic responsiveness, endothelial permeability, or immune-cell recruitment. It also offers a quantitative outcome for comparing experimental treatments.
In immunology and infection studies, the assay models how immune cells are recruited during inflammation, host defense, and pathogen-induced responses. It can also be used to examine potential anti-inflammatory or anti-infective treatments by comparing how experimental conditions alter cell passage across the barrier. The controlled in vitro format helps isolate recruitment-related processes for measurement.