Generating Mouse Intestinal Enteroids

Small intestinal enteroids were generated from 4 to 8 week old wild type C57BL6 mice as previously described [51 (link)]. Enteroids were cultured in Advanced DMEM/F12 (Life Technologies 12,634) supplemented with 1x N-2 supplement (R&D Systems AR009), 1x B27 supplement (Gibco 17,504), 10 mM HEPES (Thermo Fisher Scientific15630080), 1x Glutamax (Gibco 15,630,080), 100 U/mL Penicillin-Streptomycin (Gibco 10,378,016), 50 ng/mL recombinant mouse EGF (Peprotech 315–09), 50 ng/mL recombinant murine noggin (Peprotech 250–38), and 50 ng/mL recombinant mouse r-spondin CHO-expressed (R&D Systems 7150-RS). Organoid cultures were passaged a minimum of one time prior to experimentation.

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Dougherty M.W., Kudin O., Mühlbauer M., Neu J., Gharaibeh R.Z, & Jobin C. (2020). Gut microbiota maturation during early human life induces enterocyte proliferation via microbial metabolites. BMC Microbiology, 20, 205.

Publication 2020

Advanced DMEM/F12 N-2 supplement B27 supplement HEPES Glutamax Penicillin-Streptomycin recombinant mouse EGF recombinant murine noggin

Hepes Mouse Noggin Organoid Penicillin Small intestinal Streptomycin Supplement

Corresponding Organization : University of Florida

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Variable analysis

independent variables

Mouse genotype (wild type C57BL6)

dependent variables

Characteristics of small intestinal enteroids (culture and passage)

control variables

Age of mice (4 to 8 weeks old)
Culture media composition (Advanced DMEM/F12, N-2 supplement, B27 supplement, HEPES, Glutamax, Penicillin-Streptomycin, EGF, Noggin, R-spondin)

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

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