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

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4, 290 mosmol/1 and centrifuged at 900 g for 25 min. Plasma-free platelets were recovered at the interface, resuspended in the same buffer, counted and diluted at 109/ml. Adhesion assay. 100/zl of the platelet suspension are plated into fiat-bottomed plastic microtiter wells and allowed to adhere for 24 h at 4~ Nonadherent platelets are removed by washing with PBS/I% BSA. 100/zl of this solution are added to each well and incubated for 1 h at 37~ to block 'free adherent sites' on plastic. If required, adherent platelets can now be treated with the desired reagent(s), and then rinsed again in PBS/BSA.

At the end of each time point (a good kinetics should include at least four points), dishes are rinsed three times with PBS, which is 58 CANCERMETASTASIS gently delivered on the surface through a pipette (take care in using always the same method and force in the rinsing procedure, because application of different shear forces among experiments may alter their reproducibility) with the flow directed against the wall of the dish. Next, detection and quantitation of tumor cells adherent to specific substrates can be done by either radioactive or nonradioactive methods.

However, effects of cytokine activation of EC are not only limited to the induction of adhesion molecules and paracrine growth factors (all favorable events for metastases development). , 1989). , 1991). This observation might contribute to explain the inability of certain tumor cells to metastatize in the lung, despite their ability to efficiently lodge at this site. , 1981). Organ-specifically modulated endothelial cells. Endothelial cells, similarly to most of the cells that form a living organism, are in close contact with extracellular matrix (ECM) molecules, which are known to play important roles in regulating their phenotypic expression, as clearly shown in an in vitro system by Madri and Williams (1983).

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