Broad scope method for creating humanized animal models for animal health and disease research through antibiotic treatment and human fecal transfer

Abstract

Traditionally, mouse humanization studies have used human fecal transfer to germ-free animals. This practice requires gnotobiotic facilities and is restricted to gnotobiotic mouse lines, which limits humanized mouse research. We have developed a generalizable method to humanize non germ-free mice using antibiotic treatment and human fecal transfer. The method involves depleting resident intestinal microbiota with broad-spectrum antibiotics, introducing human microbiota from frozen fecal samples by weekly gavage, and maintaining mice in HEPA-filtered microisolator cages. Pyrosequencing cecal microbiota 16S rRNA genes showed that recipient mice adopt a humanized microbiota profile analogous to their human donors, and distinct from mice treated with only antibiotics (no fecal transfer) or untreated control mice. In the humanized mice, 75% of the sequence mass was observed in their respective human donor and conversely, 68% of the donor sequence mass was recovered in the recipient mice. Principal component analyses of GC- and HPLC-separated cecal metabolites were performed to determine effects of transplanted microbiota on the metabolome. Cecal metabolite profiles of mice treated with only antibiotics (no fecal transfer) and control mice were dissimilar from each other and from humanized mice. Metabolite profiles for mice humanized from different donor samples clustered near each other, yet were sufficiently distinct that separate clusters were apparent for each donor. Also, cecal concentrations of 57 metabolites were significantly different between humanization treatments. These data demonstrate that our protocol can be used to humanize non germ-free mice and is sufficiently robust to generate metabolomic differences between mice humanized from different human donors.

Antibiotics-induced depletion of mice microbiota induces changes in host serotonin biosynthesis and intestinal motility

“…To determine the importance of commensal microbiota in gut motility, mice were subjected to a 4-week oral administration of antibiotics combination (ampicillin, neomycin sulfate, metronidazole, and vancomycin) [17, 18, 20, 28, 29]. The antibiotics resulted in changes in the composition of commensal bacteria examined by 16S rRNA analysis.…”

“…To the best of our knowledge, the germ-free mice model has several limitations as germ-free mice are born in aseptic conditions, which may not only affect the development of the gut motility system, but also the immunity system and metabolic function [16]. In some studies, broad-spectrum antibiotics were used to deplete gut commensal microflora for creating pseudo-germ-free mice [17–20]. The antibiotics-treated method has been used to prove the reversibility of microbial effects on host 5-HT and gut motility [12].…”

In vitro vascularized liver and tumor tissue microenvironments on a chip for dynamic determination of nanoparticle transport and toxicity

“…B. Kim, ; Rongvaux et al, ; Seok et al, ). Furthermore, animals and humans have different physiology, and recent studies report poor correlation between in vivo results from animal experiments and drug efficacy and toxicity data from humans (Engelman & Kerr, ; Fernandes & Vanbever, ; Hintze et al, ; Kang & Kim, ; J. B. Kim, ; Rongvaux et al, ; Seok et al, ).…”

Improving natural product research translation: From source to clinical trial

“…Specimen collection, analysis, and archiving. [178][179][180] Inbred mice reconstituted with microbiota from wild mice have also been reported to have increased translational value. In analyses, include approaches to control for known artefacts, such as reagent contamination that can strongly affect outcomes in low biomass microbiome specimens.…”