DMEM–Dulbecco's Modified Eagle Medium

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	supplemented with 10% fetal calf serum (FBS; Invitrogen) and 1% antibiotic/antimycotic (AA, GIBCO).

Protocol tips
supplemented with 10% fetal calf serum (FBS; Invitrogen) and 1% antibiotic/antimycotic (AA, GIBCO).

Publication protocol

Cell culture
MDCK II cells were cultured in Dulbecco’s modified Eagle’s medium (Invitrogen) supplemented with 10% fetal calf serum (FBS; Invitrogen) and 1% antibiotic/antimycotic (AA, GIBCO). Cells were maintained at 37°C in a humidified atmosphere with 5% CO2, and the medium was refreshed once every 2 days.

Three-dimensional culture
MDCK II cells were added to the ice-cold DMEM containing 2 mg/ml PureCol purified collagen I (INAMED Biomaterials Cat: 5005-B), 20 mM HEPES (pH 7.6), 12 mM NaOH and 10% FBS (final concentration: 60000 cells/ml). The resulting suspension was placed into 24-well plate or Transwell chambers. After formation of a gel at 37°C, The growth medium was added to each well. Then, cells were incubated for several days for cyst formation on the gel.

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