Murine Enteroid Derivation and Maintenance

Murine enteroids were derived from C57BL/6 mice and maintained as previously described [24 (link)]. Briefly, established enteroids were maintained by embedding them in 50 μl Matrigel (Corning) and covering the domes with Mouse IntestiCult (Stemcell Technologies) 1×Pen-Strep (Corning), incubated at 37°C and 5% CO₂. Medium was replaced every 2 to 4 days and enteroids were subcultured every 5 to 7 days by mechanical sheering in Gentle Cell Dissociation Reagent (Stemcell Technologies), washing in DMEM/F-12 15 mM HEPES (Stemcell Technologies) and re-embedding in Matrigel at a 1:3 to 1:4 splitting ratio.

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Ernst C., Andreassen P.R., Giger G.H., Nguyen B.D., Gäbelein C.G., Guillaume-Gentil O., Fattinger S.A., Sellin M.E., Hardt W.D, & Vorholt J.A. (2024). Direct Salmonella injection into enteroid cells allows the study of host–pathogen interactions in the cytosol with high spatiotemporal resolution. PLOS Biology, 22(4), e3002597.

Publication 2024

Matrigel Mouse IntestiCult ×Pen-Strep Gentle Cell Dissociation Reagent

Corresponding Organization : Uppsala University

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Variable analysis

independent variables

Murine enteroids derived from C57BL/6 mice

dependent variables

Outcomes of the enteroid cultures

control variables

Matrigel (Corning) used for embedding the enteroids
Mouse IntestiCult (Stemcell Technologies) 1×Pen-Strep (Corning) used as the culture medium
Incubation conditions (37°C and 5% CO2)
Medium replacement every 2 to 4 days
Enteroid subculturing every 5 to 7 days by mechanical sheering in Gentle Cell Dissociation Reagent (Stemcell Technologies), washing in DMEM/F-12 15 mM HEPES (Stemcell Technologies) and re-embedding in Matrigel at a 1:3 to 1:4 splitting ratio

controls

No positive or negative controls were explicitly mentioned in the provided information.

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