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Automated continuous monitoring of growth factor-mediated Endothelial cell migration

Dec 5, 2017

The integrated nature of cell migration is exemplified by angiogenesis. Angiogenesis or neo-angiogenesis refers to the formation of new blood vessels from pre-existing vessels and is critical for development, wound healing and tumor growth. Endothelial cell migration is an important component of angiogenesis, involving chemotactic, haptotactic and mechanotactic (shear stress) induced cell migration. Chemotactic cell migration is typically induced by soluble growth factors such as vascular endothelial growth factor (VEGF) and its isoforms, fibroblast growth factor (bFGF) and hepatocyte growth factor (HGF) amongst others. These growth factors interact with their cognate receptor tyrosine kinases on he surface of endothelial cells activating signaling pathways culminating in directed cell migration.

In the present study, we used the CIM-Plate 16 with the xCELLigence RTCA DP Instrument to monitor growth factor-mediated migration of endothelial cells in realtime using label-free conditions. The CIM-Plate 16 is a 16-well modified Boyden chamber composed of an upper chamber (UC) and a lower chamber (LC). The UC and LC easily snap together to form a tight seal. The UC is sealed at its bottom by a microporous Polyethylene terephthalate (PET) mem-brane. These micropores permit the physical translocation of cells from the upper part of the UC to the bottom side of the membrane. The bottom side of the membrane (the side facing the LC) contains interdigitated gold microelectrode sensors which will come in contact with migrated cells and generate an impedance signal. The LC contains 16 wells, each of which serves as a reservoir for a chemoattractant solution on the bottom side of the wells, separated from each other by pressure-sensitive O-ring seals.

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Agilent technologies

Agilent provides xCELLigence impedance-based, label-free, real time cell analysis system and NovoCyte flow cytometers.

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Located inside a tissue culture incubator, the RTCA DP Analyzer has three integrated stations for E-Plates 16 or CIM-Plates 16

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