Gut 3 Gel: A High Throughput Mucus Model for Culturing Human Intestinal Microbiota

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Abstract
The human intestinal microbiota plays a crucial role in health and disease, yet recreating its complex interactions in vitro remains a significant challenge. Gut 3 Gel introduced herein as a novel in vitro mucus model, designed for culturing complex microbial communities without the need for anaerobic conditions. Intestinal microbiota samples from five donors were individually inoculated in Gut 3 Gel and cultured for 72 hours. Taxonomic composition assessment revealed that Gut 3 Gel sustains diverse microbial species and particularly promotes the growth of mucus-associated bacteria including Bifidobacterium , Lactobacillus , and Faecalibacterium . Microbial metabolic activity within Gut 3 Gel was confirmed by the increased production of acetate and butyrate, as well as of exopolysaccharides. Gut 3 Gel reproduces physiological features of intestinal mucus, providing a reproducible and scalable culturing platform. These features make Gut 3 Gel a promising tool for advancing microbiota research with potential applications in drug screening, microbiome mining, and high throughput testing of microbiome-modulating molecules.

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