A simple coculture system shows mutualism between anaerobic faecalibacteria and epithelial Caco-2 cells.

ABSTRACT: Most gut bacteria are obligate anaerobes and are important for human health. However, little mechanistic insight is available on the health benefits of specific anaerobic gut bacteria. A main obstacle in generating such knowledge is the lack of simple and robust coculturing methods for anaerobic bacteria and oxygen-requiring human cells. Here, we describe the development of a coculture system for intestinal Caco-2 cells and an anaerobic symbiont, Faecalibacterium prausnitzii, making use of 50?mL culture tubes. F. prausnitzii was grown in 40?mL YCFAG-agar with glass-adhered Caco-2 cells placed on top in 10?mL DMEM medium. Grown for 18-36?h in a humidified incubator at 37?°C and 5% CO2, coverslip-attached Caco-2 cells promoted growth and metabolism of F. prausnitzii, while F. prausnitzii suppressed inflammation and oxidative stress in Caco-2 cells. F. prausnitzii did not compromise Caco-2 cell viability. Exogenously added porcine mucin also promoted growth of F. prausnitzii, suggesting that it may be part of the mechanism of Caco-2-stimulated growth of F. prausnitzii. This 'Human oxygen-Bacteria anaerobic' (HoxBan) coculturing system uniquely establishes host-microbe mutualism of a beneficial anaerobic gut microbe in vitro and principally allows the analysis of host-microbe interactions of pure and mixed cultures of bacteria and human cells.

SUBMITTER: Sadaghian Sadabad M 

PROVIDER: S-EPMC4678368 | biostudies-literature | 2015 Dec

REPOSITORIES: biostudies-literature