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Merck

Adhesion and Invasion Assay Procedure Using Caco-2 Cells for Listeria monocytogenes.

Bio-protocol (2017-05-05)
Swetha Reddy, Frank Austin
RESUMEN

Listeria monocytogenes is an important Gram-positive foodborne pathogen that is a particular problem in ready-to-eat food. It has an ability to survive in harsh conditions like refrigeration temperatures and high salt concentrations and is known to cross intestinal, placental and blood-brain barriers. Several cancerous cell lines like cervical, liver, dendritic, intestinal and macrophages have been used to study in vitro propagation and survival of listeria in human cells. Human intestinal epithelial cells have been used to study how listeria crosses the intestinal barrier and cause infection. The protocol in this articles describes the procedures to grow Caco-2 cells, maintain cells and use them for adhesion and invasion assays. During adhesion assay the cells are incubated with listeria for 30 min but in invasion assay the cell growth is arrested at several time points after infection to monitor the growth and survival rate of listeria in cells.

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Sigma-Aldrich
Disolución salina tamponada con fosfatos de Dulbecco, Modified, without calcium chloride and magnesium chloride, liquid, sterile-filtered, suitable for cell culture
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Gentamicina solution, 50 mg/mL in deionized water, liquid, 0.1 μm filtered, BioReagent, suitable for cell culture
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Disolución salina tamponada con fosfato, 10× concentrate, BioPerformance Certified, suitable for cell culture