Ulluwishewa Dulantha, Anderson Rachel C, Young Wayne, McNabb Warren C, van Baarlen Peter, Moughan Paul J, Wells Jerry M, Roy Nicole C
Food Nutrition & Health Team, Food & Bio-Based Products Group, AgResearch Grasslands, Palmerston North, 4442, New Zealand; Riddet Institute, Massey University, Palmerston North, 4442, New Zealand.
Cell Microbiol. 2015 Feb;17(2):226-40. doi: 10.1111/cmi.12360. Epub 2014 Oct 31.
Faecalibacterium prausnitzii, an abundant member of the human commensal microbiota, has been proposed to have a protective role in the intestine. However, it is an obligate anaerobe, difficult to co-culture in viable form with oxygen-requiring intestinal cells. To overcome this limitation, a unique apical anaerobic model of the intestinal barrier, which enabled co-culture of live obligate anaerobes with the human intestinal cell line Caco-2, was developed. Caco-2 cells remained viable and maintained an intact barrier for at least 12 h, consistent with gene expression data, which suggested Caco-2 cells had adapted to survive in an oxygen-reduced atmosphere. Live F. prausnitzii cells, but not ultraviolet (UV)-killed F. prausnitzii, increased the permeability of mannitol across the epithelial barrier. Gene expression analysis showed inflammatory mediators to be expressed at lower amounts in Caco-2 cells exposed to live F. prausnitzii than UV-killed F. prausnitzii, This, consistent with previous reports, implies that live F. prausnitzii produces an anti-inflammatory compound in the culture supernatant, demonstrating the value of a physiologically relevant co-culture system that allows obligate anaerobic bacteria to remain viable.
普拉梭菌是人类共生微生物群中数量丰富的一员，被认为在肠道中具有保护作用。然而，它是一种专性厌氧菌，很难与需氧肠道细胞以活的形式共培养。为了克服这一限制，开发了一种独特的肠道屏障顶端厌氧模型，该模型能够使活的专性厌氧菌与人肠道细胞系Caco-2共培养。Caco-2细胞保持存活并维持完整屏障至少12小时，这与基因表达数据一致，表明Caco-2细胞已适应在低氧环境中存活。活的普拉梭菌细胞，而不是紫外线（UV）杀死的普拉梭菌，增加了甘露醇跨上皮屏障的通透性。基因表达分析表明，与紫外线杀死的普拉梭菌相比，暴露于活的普拉梭菌的Caco-2细胞中炎症介质的表达量更低。这与之前的报道一致，意味着活的普拉梭菌在培养上清液中产生一种抗炎化合物，证明了一个生理相关的共培养系统的价值，该系统允许专性厌氧菌保持存活。
