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Interaction of Rat Ascites Hepatoma Cells with Cultured Mesothelial Cell Layers: A Model for Tumor Invasion¹

Departments of Tumor Biochemistry [H. A., K. S., M. M.], Cell Biology [Y. M.], Experimental Pathology [R. T., K. T.], Gastrointestinal Oncology [R. Y.], Research Institute, and Department of Clinical Cytology [T. M.], Hosptial, The Center for Adult Diseases, Osaka, Osaka 537, Japan

Interactions of rat ascites hepatoma cells with primary cultured layers of rat mesentery-derived cells were studied. The mesentery-derived cells were isolated from rat mesentery and cultured in Eagle's minimum essential medium with a 2-fold concentration of amino acids and vitamins supplemented with 10% calf serum. The primary cultured cells, consisting mainly of mesothelial cells in polygonal shape, forms a "paving stone" sheet. Upon seeding the tumor cells on the mesentery-derived cell alyers, three different types of tumor cell growth were observed. Type 1 was the formation of piled-up tumor cell nests on mesothelial cell layers. Type 2 was the formation of flattened tumor cell islands underneath mesothelial cell layers. This island formation was clearly observed under a phase contrast microscope 2 days after the tumor cell seeding. Protrusion of cellular processes of the tumor cells beneath mesothelial cells was occasionally seen. Type 3 was the growth of tumor cells in suspension. These types of tumor cell growth closely resemble those in the peritoneal cavity observed after i.p. implantation of the tumor cells. When the tumor cells recovered from the blood of tumor-bearing rats were seeded, flattened tumor cell islands were formed 15 times more frequently than when the tumor cells isolated from host peritoneal cavity were seeded. Shortly after the appearance of small flattened tumor cell islands, a distinct morphological change of mesothelial cells from polygonal to spindle shape was seen preferentially at the marginal area of the cell layers (a partial retraction of cell edges). The retraction of mesothelial cells was induced not only by seeding the tumor cells but by adding the tumor ascites fluid or the medium conditioned by the tumor cell culture. The morphological change was reversed by changing the culture medium to remove the effectors. These results indicate that the system described in this study can provide a useful model to study tumor cell invasion.

¹ Supported in part by a Grant-in-Aid for Cancer Research from the Japanese Ministry of Education and by the Osaka Association for the Prevention of Adult Diseases.

² To whom requests for reprints should be addressed.

Received 1/29/85. Revised 10/ 4/85. Revised 1/15/86. Accepted 1/16/86.

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