MDCK cells were purchased from Riken Cell Bank and cultured in Dulbecco's modified Eagle's medium (DMEM; Sigma, St. Louis, MO) containing 1% antibiotic/antimycotic solution (Sigma) and 10% fetal bovine serum (FBS; Equitech-Bio, Inc., Kerrville, TX). Cells were maintained in a humidified incubator at 37°C and 5% CO2. For time-lapse imaging, immunofluorescence staining, and live cell imaging, we prepared a 1.6 mg/ml collagen type I gel with Cellmatrix type I-P (Nitta Gelatin Inc., Osaka, Japan). To form a circular monolayer colony, a hand-made plastic ring (diameter, 5 mm) was placed on a collagen gel. The cell suspension obtained after trypsinization was dropped within the ring and incubated for 6 h. The ring was removed, and unattached cells were washed away (Supplementary Fig. 8a ). After 2–3 days, several leader cells were observed in each dish.
Mdck cells
Manufactured by RIKEN Cell Bank
Sourced in Japan
MDCK cells are a well-established epithelial cell line derived from the kidney of a normal adult female cocker spaniel. These cells are widely used as a model system for studying various aspects of epithelial cell biology, including cell-cell interactions, transport processes, and signal transduction pathways.
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2 protocols using mdck cells
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MDCK Cell Monolayer Formation on Collagen Gel
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Culturing MDCK Cells in DMEM
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MDCK cells from Riken Cell Bank (Tsukuba, Japan) were cultured in DMEM supplemented with 10% fetal calf serum (FCS), 1% penicillin-streptomycin (Nacalai Tesque), and 2 mM glutamine (FCS-DMEM). Cells were cultivated in a humidified atmosphere containing 5% CO2 at 37 °C.
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