title: 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.
authors.affiliation: Departments of Medical Microbiology, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands.|Departments of Gastroenterology and Hepatology, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands.
authors.forename: Mehdi
authors.initials: M
authors.lastname: Sadaghian Sadabad
authors.affiliation: Departments of Gastroenterology and Hepatology, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands.
authors.forename: Julius Z H
authors.initials: JZ
authors.lastname: von Martels
authors.affiliation: Departments of Medical Microbiology, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands.
authors.forename: Muhammed Tanweer
authors.initials: MT
authors.lastname: Khan
authors.affiliation: Departments of Laboratory Medicine, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands.
authors.forename: Tjasso
authors.initials: T
authors.lastname: Blokzijl
authors.affiliation: Istituto Zooprofilattico Sperimentale di Puglia e Basilicata, Foggia, Italy.|Center for Systems Biology University of Iceland Reykjavik, Iceland.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Paglia
authors.affiliation: Departments of Gastroenterology and Hepatology, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands.
authors.forename: Gerard
authors.initials: G
authors.lastname: Dijkstra
authors.affiliation: Departments of Medical Microbiology, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands.
authors.forename: Hermie J M
authors.initials: HJ
authors.lastname: Harmsen
authors.affiliation: Departments of Gastroenterology and Hepatology, University of Groningen, University Medical Center Groningen, Groningen, The Netherlands.
authors.forename: Klaas Nico
authors.initials: KN
authors.lastname: Faber
mesh_terms: D001421:Bacteria, Anaerobic / Q000502:physiology; D015415:Biomarkers; D018938:Caco-2 Cells; D018920:Coculture Techniques; D017077:Culture Media, Conditioned / Q000378:metabolism; D004847:Epithelial Cells / Q000502:physiology; D006801:Humans; D007413:Intestinal Mucosa / Q000378:metabolism / Q000382:microbiology; D055442:Metabolome; D055432:Metabolomics / Q000379:methods; D009077:Mucins / Q000378:metabolism; D013559:Symbiosis
chemical_list: D015415:Biomarkers; D017077:Culture Media, Conditioned; D009077:Mucins