Gibco™Advanced DMEM/F-12

Protocol tips

Upstream tips	Protocol tips	Downstream tips
Defrost in advance basement membrane extract (BME; Cultrex) on ice or in fridge	Use uncoated 24-well plates Refresh medium every 2-3 days

Upstream tips
Defrost in advance basement membrane extract (BME; Cultrex) on ice or in fridge
Protocol tips
Use uncoated 24-well plates Refresh medium every 2-3 days

Publication protocol

Organoid culture: Single cells were seeded in basement membrane extract (BME; Cultrex) in uncoated 24-well plates (four drops of 10 μl per well containing 25 cells, for Confetti-fluorescent cells up to 200) and cultured in Advanced DMEM/F12 (Gibco) supplemented with factors listed in Table S1 with or without Wtn3a and R-spondin-2 conditioned medium, or FGF2 and heparin for 12-14 days. Milk production was induced by incubation with minimal medium with or without prolactin for 48-96 h. ROCK inhibitor (Y27632) was added for the first 3 days and the medium was refreshed every 2-3 days. For organoid proliferation measurements, 25 single cells/10 μl BME were plated in 96-well plates (Nunc). After 12-14 days, CellTiter-Glo Luminescent Cell Viability Assay (Promega) was used to measure cell proliferation. For flow cytometry, 12- to 14-day-old organoids were dissociated into single cells using TrypLE express (Thermo Fisher Scientific) prior to staining.

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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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