Gibco™Advanced DMEM/F-12

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Defrost the Matrigel in advance on ice or in fridge	Use of Matrigel or a collagen-based matrix

Upstream tips
Defrost the Matrigel in advance on ice or in fridge
Protocol tips
Use of Matrigel or a collagen-based matrix

Publication protocol

Isolation and Culture of Mouse Stomach Organoid: Mouse stomach tissue was obtained from the pyloric antrum to the fundus. The tissue was then opened and washed several times in ice-cold DPBS. To isolate the gastric glands, the tissue was put into 5 ml of ice-cold chelating buffer (5 mM EDTA in DPBS) and incubated for 2 hours on an orbital shaker at room temperature. After incubation, 5 ml of dissociation buffer was added and gently inverted by hand for 1–2 minutes. Next, the gastric glands were filtered through a cell strainer and plated with Matrigel or a collagen-based matrix at a ratio of 1 : 1 and seeded in multiwell plates. After polymerization of the matrices, mouse stomach organoid growth medium was added. The composition of the mouse stomach organoid growth medium was as follows: basal growth media supplemented with 10% R-Spondin 1 conditioned medium, mEGF, mNoggin, 50% Wnt-3A conditioned medium, 500 nM A83-01, 10 μM Y-27632, 10 nM [Leu15]-Gastrin 1, 200 ng/ml FGF10, and 2.5 μM CHIR99021 (Table 1).

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Manufacturer protocol

Download the product protocol from Thermo Fisher Scientific for Gibco™Advanced DMEM/F-12 below.

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