Primary human colonic mucosal barrier crosstalk with super oxygen-sensitiveFaecalibacterium prausnitziiin continuous culture

Abstract

<jats:title>Abstract</jats:title><jats:p>The gut microbiome plays an important role in human health and disease. Gnotobiotic animal and <jats:italic>in vitro</jats:italic> cell-based models provide some informative insights into mechanistic crosstalk. However, there is no existing system for a chronic co-culture of a human colonic mucosal barrier with super oxygen-sensitive commensal microbes, hindering the study of human-microbe interactions in a controlled manner. Here, we investigated the effects of an abundant super oxygen-sensitive commensal anaerobe, <jats:italic>Faecalibacterium prausnitzii</jats:italic>, on a primary human mucosal barrier using a Gut-MIcrobiome (GuMI) physiome platform that we designed and fabricated. Chronic continuous co-culture of <jats:italic>F. prausnitzii</jats:italic> for two days with colon epithelia, enabled by continuous flow of completely anoxic apical media and aerobic basal media, resulted in a strictly anaerobic apical environment fostering growth of and butyrate production by <jats:italic>F. prausnitzii</jats:italic>, while maintaining a stable colon epithelial barrier. We identified elevated differentiation and hypoxia-responsive genes and pathways in the platform compared with conventional aerobic static culture of the colon epithelia, attributable to a combination of anaerobic environment and continuous medium replenishment. Furthermore, we demonstrated anti-inflammatory effects of <jats:italic>F. prausnitzii</jats:italic> through HDAC and the TLR-NFKB axis. Finally, we identified that butyrate largely contributes to the anti-inflammatory effects by downregulating TLR3 and TLR4. Our results are consistent with some clinical observations regarding <jats:italic>F. prausnitzii</jats:italic>, thus motivating further studies employing this platform with more complex engineered colon tissues for understanding the interaction between the human colonic mucosal barrier and microbiota, pathogens, or engineered bacteria.</jats:p>

Intestinal explant barrier chip: long-term intestinal absorption screening in a novel microphysiological system using tissue explants

“…Furthermore, development of an aerobic-anaerobic interface would facilitate a more physiological environment for the intestinal tissue, and thereby also create the possibility to incorporate living gut microbiome which has an abundance of strict anaerobes. A few examples of gut-on-a-chip models with an aerobicanaerobic interface have been presented in the past years, [80][81][82][83] but none of these systems use tissue explants and all are technically challenging to operate. Nevertheless, it would be interesting to create such an interface for the IEBC to incorporate the gut microbiome in future research.…”

Fecal luminal factors from patients with irritable bowel syndrome induce distinct gene expression of colonoids

“…The colonoid monolayers in our study presented with similar features, that is, polarization and specialized cells as mucin-producing goblet cells, to those previously described in the literature. 27,[33][34][35] The addition of F I G U R E 6 Fecal microbiota and metabolite profiles from healthy subjects and IBS patients. Fecal samples were analyzed by GAmap ™ dysbiosis test while fecal supernatants were analyzed by untargeted liquid chromatography/mass spectrometry analysis from healthy subjects (n = 7 gray dots) or IBS patients (n = 9 black dots).…”

Critical Considerations for the Design of Multi-Organ Microphysiological Systems (MPS)

“…The integral role of the microbiome in gut function and homeostasis is represented in a gut-on-a-chip model using bacteria such as Escherichia coli (Tovaglieri et al, 2019), Faecalibacterium prausnitzii (Zhang J. et al, 2021), and Lactobacillus rhamnosus and Bacteroides caccae (Shah et al, 2016). The co-culture of epithelial cells with microbes to model microbe-host cell relationships provides a better understanding of their involvement in diseased and healthy intestinal conditions.…”