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An epithelial Madin-Darby canine kidney (MDCK) cell line was grown in two kinds of standard media and a medium conditioned by MRC-5 embryonic lung cells, and temporo-regional differences in cell proliferation were examined by immunohistochemical detection of incorporated bromodeoxyuridine (BrdU). MRC-5-conditioned medium is known to contain scatter factor, which induces scattering of MDCK cells. In a proliferation assay, the BrdU labeling index (percentage ratio of BrdU-positive to total nuclei) was highest in day 2 cultures in the standard media, but on day 3 in the conditioned medium. MDCK cells in the standard media formed multiple epithelial sheets by day 2. In the conditioned medium, however, epithelial sheet formation was retarded and observed only after day 3. Once epithelial sheets had formed, cells close to the edge of the sheet were more proliferative than those distant from the edge in both standard and conditioned media. These findings suggest that proliferation of MDCK cells is suppressed during the scattering induced by the conditioned medium, and that their DNA synthesis becomes most active when cell scattering has ceased and epithelial sheets have begun to form.

The Influence of Scattering on Temporo-regional Proliferation in the Cultured Madin-Darby Canine Kidney (MDCK) Cells.