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A guide to germ-free and gnotobiotic mouse technology to study health and disease

Maude Jans (UGent) and Lars Vereecke (UGent)
(2025) FEBS JOURNAL. 292(6). p.1228-1251
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Abstract
The intestinal microbiota has major influence on human physiology and modulates health and disease. Complex host-microbe interactions regulate various homeostatic processes, including metabolism and immune function, while disturbances in microbiota composition (dysbiosis) are associated with a plethora of human diseases and are believed to modulate disease initiation, progression and therapy response. The vast complexity of the human microbiota and its metabolic output represents a great challenge in unraveling the molecular basis of host-microbe interactions in specific physiological contexts. To increase our understanding of these interactions, functional microbiota research using animal models in a reductionistic setting are essential. In the dynamic landscape of gut microbiota research, the use of germ-free and gnotobiotic mouse technology, in which causal disease-driving mechanisms can be dissected, represents a pivotal investigative tool for functional microbiota research in health and disease, in which causal disease-driving mechanisms can be dissected. A better understanding of the health-modulating functions of the microbiota opens perspectives for improved therapies in many diseases. In this review, we discuss practical considerations for the design and execution of germ-free and gnotobiotic experiments, including considerations around germ-free rederivation and housing conditions, route and timing of microbial administration, and dosing protocols. This comprehensive overview aims to provide researchers with valuable insights for improved experimental design in the field of functional microbiota research. Germ-free and gnotobiotic mouse technology allows the unraveling of complex host-microbiota interactions within a reductionist framework. The implementation of these methodologies poses significant technical and experimental challenges. This review explores practical insights aimed at enhancing the precision and success of germ-free and gnotobiotic experiments. image
Keywords
microbiota, gnotobiotic, germ-free, experimental design, ALTERED SCHAEDLER FLORA, SEGMENTED FILAMENTOUS BACTERIA, ARYL-HYDROCARBON RECEPTOR, GUT MICROBIOTA PROTECTS, FREE-MICE, INTESTINAL INFLAMMATION, ULCERATIVE-COLITIS, COMMENSAL BACTERIA, DEVELOP COLITIS, PIG MODEL

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Citation

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MLA
Jans, Maude, and Lars Vereecke. “A Guide to Germ-Free and Gnotobiotic Mouse Technology to Study Health and Disease.” FEBS JOURNAL, vol. 292, no. 6, 2025, pp. 1228–51, doi:10.1111/febs.17124.
APA
Jans, M., & Vereecke, L. (2025). A guide to germ-free and gnotobiotic mouse technology to study health and disease. FEBS JOURNAL, 292(6), 1228–1251. https://doi.org/10.1111/febs.17124
Chicago author-date
Jans, Maude, and Lars Vereecke. 2025. “A Guide to Germ-Free and Gnotobiotic Mouse Technology to Study Health and Disease.” FEBS JOURNAL 292 (6): 1228–51. https://doi.org/10.1111/febs.17124.
Chicago author-date (all authors)
Jans, Maude, and Lars Vereecke. 2025. “A Guide to Germ-Free and Gnotobiotic Mouse Technology to Study Health and Disease.” FEBS JOURNAL 292 (6): 1228–1251. doi:10.1111/febs.17124.
Vancouver
1.
Jans M, Vereecke L. A guide to germ-free and gnotobiotic mouse technology to study health and disease. FEBS JOURNAL. 2025;292(6):1228–51.
IEEE
[1]
M. Jans and L. Vereecke, “A guide to germ-free and gnotobiotic mouse technology to study health and disease,” FEBS JOURNAL, vol. 292, no. 6, pp. 1228–1251, 2025.
@article{01HY3C3PZC75DMHTQAC52FK1JE,
  abstract     = {{The intestinal microbiota has major influence on human physiology and modulates health and disease. Complex host-microbe interactions regulate various homeostatic processes, including metabolism and immune function, while disturbances in microbiota composition (dysbiosis) are associated with a plethora of human diseases and are believed to modulate disease initiation, progression and therapy response. The vast complexity of the human microbiota and its metabolic output represents a great challenge in unraveling the molecular basis of host-microbe interactions in specific physiological contexts. To increase our understanding of these interactions, functional microbiota research using animal models in a reductionistic setting are essential. In the dynamic landscape of gut microbiota research, the use of germ-free and gnotobiotic mouse technology, in which causal disease-driving mechanisms can be dissected, represents a pivotal investigative tool for functional microbiota research in health and disease, in which causal disease-driving mechanisms can be dissected. A better understanding of the health-modulating functions of the microbiota opens perspectives for improved therapies in many diseases. In this review, we discuss practical considerations for the design and execution of germ-free and gnotobiotic experiments, including considerations around germ-free rederivation and housing conditions, route and timing of microbial administration, and dosing protocols. This comprehensive overview aims to provide researchers with valuable insights for improved experimental design in the field of functional microbiota research.

 Germ-free and gnotobiotic mouse technology allows the unraveling of complex host-microbiota interactions within a reductionist framework. The implementation of these methodologies poses significant technical and experimental challenges. This review explores practical insights aimed at enhancing the precision and success of germ-free and gnotobiotic experiments. image}},
  author       = {{Jans, Maude and Vereecke, Lars}},
  issn         = {{1742-464X}},
  journal      = {{FEBS JOURNAL}},
  keywords     = {{microbiota,gnotobiotic,germ-free,experimental design,ALTERED SCHAEDLER FLORA,SEGMENTED FILAMENTOUS BACTERIA,ARYL-HYDROCARBON RECEPTOR,GUT MICROBIOTA PROTECTS,FREE-MICE,INTESTINAL INFLAMMATION,ULCERATIVE-COLITIS,COMMENSAL BACTERIA,DEVELOP COLITIS,PIG MODEL}},
  language     = {{eng}},
  number       = {{6}},
  pages        = {{1228--1251}},
  title        = {{A guide to germ-free and gnotobiotic mouse technology to study health and disease}},
  url          = {{http://doi.org/10.1111/febs.17124}},
  volume       = {{292}},
  year         = {{2025}},
}

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