Abstract
Mice transplanted with human microbiota are essential tools for studying the role of microbiota in health and disease, striving for the development of microbiota-modulating therapeutics. Traditionally, germ-free mice have been the principal option for establishing human microbiota-associated (HMA) mouse models, leading to significant insights into the composition and function of the human microbiota. However, there are limitations in using germ-free mice as recipients of human microbiota, including considerable resource allocation to establish and maintain the model and incomplete development of their immune system and physiological functions. Thus, antibiotic-treated, non-germ-free mice have been developed as an alternative to satisfy the growing demand for an accessible HMA mouse model. Several methods have been described for creating “humanized” mice. These protocols vary in their key components, mainly antibiotic conditioning and frequency of oral gavage. To address this practical challenge and formulate a simple and repeatable protocol, we established a HMA mouse model with antibiotic-treated conventional and specific-pathogen free (SPF) C57BL/6J mice, revealing that a single oral gavage allows stable engraftment of the human microbiota. In this chapter, we present our simple protocol for antibiotic conditioning to prepare mice for stable engraftment of human gut microbiota.
Key words
- Antibiotics
- Dysbiosis
- Fecal microbiota transplantation
- Gut microbiota
- Human microbiota-associated mice
- Humanized mice
- Mouse model
- Oral gavage
This is a preview of subscription content, access via your institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsReferences
Ley RE, Peterson DA, Gordon JI (2006) Ecological and evolutionary forces shaping microbial diversity in the human intestine. Cell 124:837–848
Qin J, Li R, Raes J, Arumugam M, Burgdorf KS, Manichanh C et al (2010) A human gut microbial gene catalogue established by metagenomic sequencing. Nature 464:59–65
Bäckhed F, Ley RE, Sonnenburg JL, Peterson DA, Gordon JI (2005) Host-bacterial mutualism in the human intestine. Science 307:1915–1920
Chung H, Pamp SJ, Hill JA, Surana NK, Edelman SM, Troy EB et al (2012) Gut immune maturation depends on colonization with a host-specific microbiota. Cell 149:1578–1593
Lu J, Lu L, Yu Y, Cluette-Brown J, Martin CR, Claud EC (2018) Effects of intestinal microbiota on brain development in humanized gnotobiotic mice. Sci Rep 8:1–16
Dupont AW, Dupont HL (2011) The intestinal microbiota and chronic disorders of the gut. Nat Rev Gastroenterol Hepatol 8:523–531
Aron-Wisnewsky J, Prifti E, Belda E, Ichou F, Kayser BD, Dao MC et al (2019) Major microbiota dysbiosis in severe obesity: fate after bariatric surgery. Gut 68:70–82
Li J, Zhao F, Wang Y, Chen J, Tao J, Tian G et al (2017) Gut microbiota dysbiosis contributes to the development of hypertension. Microbiome 5:14
Zou S, Fang L, Lee M-H (2018) Dysbiosis of gut microbiota in promoting the development of colorectal cancer. Gastroenterol Rep 6:1–12
Sharon G, Cruz NJ, Kang DW, Gandal MJ, Wang B, Kim YM et al (2019) Human gut microbiota from autism spectrum disorder promote behavioral symptoms in mice. Cell 177:1600–1618.e17
Ridaura VK, Faith JJ, Rey FE, Cheng J, Duncan AE, Kau AL et al (2013) Gut microbiota from twins discordant for obesity modulate metabolism in mice. Science 341:1241214
Nagao-Kitamoto H, Shreiner AB, Gillilland MG, Kitamoto S, Ishii C, Hirayama A et al (2016) Functional characterization of inflammatory bowel disease-associated gut dysbiosis in gnotobiotic mice. Cell Mol Gastron Hepatol 2:468–481
Sobhani I, Bergsten E, Couffin S, Amiot A, Nebbad B, Barau C et al (2019) Colorectal cancer-associated microbiota contributes to oncogenic epigenetic signatures. Proc Natl Acad Sci U S A 116:24285–24295
Wong SH, Zhao L, Zhang X, Nakatsu G, Han J, Xu W et al (2017) Gavage of fecal samples from patients with colorectal cancer promotes intestinal carcinogenesis in germ-free and conventional mice. Gastroenterology 153:1621–1633.e6
Bäckhed F, Manchester JK, Semenkovich CF, Gordon JI (2007) Mechanisms underlying the resistance to diet-induced obesity in germ-free mice. Proc Natl Acad Sci U S A 104:979–984
Turnbaugh PJ, Ley RE, Mahowald MA, Magrini V, Mardis ER, Gordon JI (2006) An obesity-associated gut microbiome with increased capacity for energy harvest. Nature 444:1027–1031
Bäckhed F, Ding H, Wang T, Hooper LV, Koh GY, Nagy A et al (2004) The gut microbiota as an environmental factor that regulates fat storage. Proc Natl Acad Sci U S A 101:15718–15723
Hintze KJ, Cox JE, Rrompato G, Benninghoff AD, Ward RE, Broadbent J et al (2014) Broad scope method for creating humanized animal models for animal health and disease research through antibiotic treatment and human fecal transfer. Gut Microbes 5:37–41
Lundberg R, Toft MF, Metzdorff SB, Hansen CHF, Licht TR, Bahl MI et al (2020) Human microbiota-transplanted C57BL/6 mice and offspring display reduced establishment of key bacteria and reduced immune stimulation compared to mouse microbiota-transplantation. Sci Rep 10:1–16
Rodriguez J, Hiel S, Neyrinck AM, Le Roy T, Pötgens SA, Leyrolle Q et al (2020) Discovery of the gut microbial signature driving the efficacy of prebiotic intervention in obese patients. Gut:1–13
Staley C, Kaiser T, Beura LK, Hamilton MJ, Weingarden AR, Bobr A et al (2017) Stable engraftment of human microbiota into mice with a single oral gavage following antibiotic conditioning. Microbiome 5:87
Manichanh C, Reeder J, Gibert P, Varela E, Llopis M, Antolin M et al (2010) Reshaping the gut microbiome with bacterial transplantation and antibiotic intake. Genome Res 20:1411–1419
Wos-Oxley M, Bleich A, Oxley AP, Kahl S, Janus LM, Smoczek A et al (2012) Comparative evaluation of establishing a human gut microbial community within rodent models. Gut Microbes 3:234–249
Wrzosek L, Ciocan D, Borentain P, Spatz M, Puchois V, Hugot C et al (2018) Transplantation of human microbiota into conventional mice durably reshapes the gut microbiota. Sci Rep 8:6854
Nguyen TLA, Vieira-Silva S, Liston A, Raes J (2015) How informative is the mouse for human gut microbiota research? Dis Model Mech 8:1–16
Knights D, Kuczynski J, Charlson ES, Zaneveld J, Mozer MC, Collman RG et al (2011) Bayesian community-wide culture-independent microbial source tracking. Nat Methods 8:761–U107
Caporaso JG, Lauber CL, Walters WA, Berg-Lyons D, Huntley J, Fierer N et al (2012) Ultra-high-throughput microbial community analysis on the Illumina HiSeq and MiSeq platforms. ISME J 6:1621–1624
Schloss PD, Westcott SL, Ryabin T, Hall JR, Hartmann M, Hollister EB et al (2009) Introducing mothur: open-source, platform-independent, community-supported software for describing and comparing microbial communities. Appl Environ Microbiol 75:7537–7541
Pruesse E, Quast C, Knittel K, Fuchs BM, Ludwig W, Peplies J et al (2007) SILVA: a comprehensive online resource for quality checked and aligned ribosomal RNA sequence data compatible with ARB. Nucleic Acids Res 35:7188–7196
Cole JR, Wang Q, Cardenas E, Fish J, Chai B, Farris RJ et al (2009) The ribosomal database project: improved alignments and new tools for rRNA analysis. Nucleic Acids Res 37:D141–D145
Staley C, Kaiser T, Vaughn BP, Graiziger C, Hamilton MJ, Kabage AJ et al (2019) Durable long-term bacterial engraftment following encapsulated fecal microbiota transplantation to treat Clostridium difficile infection. MBio 10:e01586–e01519
Acknowledgments
We would like to thank Dr. Harika Nalluri for her input and editing of the final draft of this chapter.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2021 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Zhu, Z., Kaiser, T., Staley, C. (2021). Antibiotic Conditioning and Single Gavage Allows Stable Engraftment of Human Microbiota in Mice. In: Adami, G.R. (eds) The Oral Microbiome. Methods in Molecular Biology, vol 2327. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1518-8_17
Download citation
DOI: https://doi.org/10.1007/978-1-0716-1518-8_17
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-1517-1
Online ISBN: 978-1-0716-1518-8
eBook Packages: Springer Protocols