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  • Constructing a gnotobiotic mouse model with a synthetic human gut microbiome to study host-microbe cross talk.

Constructing a gnotobiotic mouse model with a synthetic human gut microbiome to study host-microbe cross talk.

STAR protocols (2021-06-29)
Alex Steimle, Alessandro De Sciscio, Mareike Neumann, Erica T Grant, Gabriel V Pereira, Hiroshi Ohno, Eric C Martens, Mahesh S Desai
ABSTRACT

Reproducible in vivo models are necessary to address functional aspects of the gut microbiome in various diseases. Here, we present a gnotobiotic mouse model that allows for the investigation of specific microbial functions within the microbiome. We describe how to culture 14 different well-characterized human gut species and how to verify their proper colonization in germ-free mice. This protocol can be modified to add or remove certain species of interest to investigate microbial mechanistic details in various disease models. For complete details on the use and execution of this protocol, please refer to Desai et al. (2016).

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
N-Acetyl-D-glucosamine, suitable for cell culture, BioReagent
Sigma-Aldrich
Vitamin B12, ≥98%
Sigma-Aldrich
Hemin, BioXtra, from Porcine, ≥96.0% (HPLC)
Sigma-Aldrich
Thiamine hydrochloride, reagent grade, ≥99% (HPLC)
Sigma-Aldrich
(−)-Riboflavin, BioReagent, suitable for cell culture, suitable for insect cell culture, ≥98%
Sigma-Aldrich
D-(+)-Glucose, ≥99.5% (GC), BioXtra
Sigma-Aldrich
Resazurin sodium salt, certified by the BSC
Sigma-Aldrich
Folic acid, BioReagent, suitable for cell culture, suitable for insect cell culture, suitable for plant cell culture, ≥97%
Sigma-Aldrich
Biotin, ≥99% (HPLC), lyophilized powder
Sigma-Aldrich
Phenol:Chloroform:Isoamyl Alcohol 25:24:1, Saturated with 10mM Tris, pH 8.0, 1mM EDTA, Molecular Biology
Sigma-Aldrich
Isovaleric acid, 99%
Sigma-Aldrich
Valeric acid, ≥99%