Cancer Metastasis: In vitro and in vivo Experimental by D. Rusciano, D.R. Welch and M.M. Burger (Eds.)

By D. Rusciano, D.R. Welch and M.M. Burger (Eds.)

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Once the required adhesion molecule(s) has been put in solution as recommended by the manufacturer (usually a stock solution of 1 mg/ml is prepared), a final dilution is made in sterile PBS so to evenly distribute between 1 54 CANCER METASTASIS and 10/zg/cm 2 per dish. , 1980). v. , 1992). 2 shows an adhesion assay on both adhesive (fibronectin) and nonadhesive (tenascin) substrate. Subendothelial ECM. , 1979), that can be used as a substrate to test the adhesiveness of tumor cells to a complex matrix.

1983). 5% FCS. Cells floating in the medium are aspirated. At no time cells are pipetted up and down. This gentle washing process is repeated four times. Once the matrix is free of cells, plates are rinsed with PBS at least three times. Lyric removal of EC is usually performed in two steps. 05% BSA can be added) 15 min at 37~ and then with detergent (either Ch. 2 HOMOTYPIC AND HETEROTYPICCELLADHESIONIN METASTASIS 55 Fig. 2. Positive and negative adhesion of fibroblasts on fibronectin and tenascin coating of a tissue culture dish, after saturation with BSA.

Coating with purified molecules. Once the required adhesion molecule(s) has been put in solution as recommended by the manufacturer (usually a stock solution of 1 mg/ml is prepared), a final dilution is made in sterile PBS so to evenly distribute between 1 54 CANCER METASTASIS and 10/zg/cm 2 per dish. , 1980). v. , 1992). 2 shows an adhesion assay on both adhesive (fibronectin) and nonadhesive (tenascin) substrate. Subendothelial ECM. , 1979), that can be used as a substrate to test the adhesiveness of tumor cells to a complex matrix.

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