Reddy Swetha, Austin Frank
College of Veterinary Science, Mississippi State University, Starkville, USA.
Bio Protoc. 2017 May 5;7(9):e2267. doi: 10.21769/BioProtoc.2267.
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 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.
是一种重要的革兰氏阳性食源性病原体，在即食食品中是一个特别突出的问题。它有能力在诸如冷藏温度和高盐浓度等恶劣条件下存活，并且已知能穿过肠道、胎盘和血脑屏障。几种癌细胞系，如宫颈、肝脏、树突状、肠道和巨噬细胞系，已被用于研究李斯特菌在人体细胞中的繁殖和存活情况。人类肠道上皮细胞已被用于研究李斯特菌如何穿过肠道屏障并引发感染。本文中的实验方案描述了培养Caco-2细胞、维持细胞状态并将其用于黏附与侵袭试验的步骤。在黏附试验中，细胞与李斯特菌一起孵育30分钟，但在侵袭试验中，在感染后的几个时间点阻断细胞生长，以监测李斯特菌在细胞中的生长和存活率。
