The principal goals of the proposed experiments are (1) to evaluate the influence of intracellular communications (requiring direct cell contact) or intercellular interactions (e.g., diffusible factors) between normal and carcinogen altered cells """"""""cocultured"""""""" in vivo, (2) to evaluate whether carcinogen altered cells lose the ability to be influenced by or influence the behavior of, other populations in vivo as they progress from a normal to a neoplastic state; and (3) to determine whether TPA affects those cellular interactions which normally inhibit development and expression of the neoplastic phenotype in the intact tissue. Suspensions of normal and carcinogen altered cells or pairs of different carcinogen-altered cells will be mixed in ratios varying from 1:1 to 1:100 and cocultured in vivo in denuded tracheal grafts. At various times up to 1 year after exposure cells will be harvested enzymatically, populations quantitated and separated by flow cytometry and cell culture techniques. The effect of cell mixing ratios, and spatial configuration of mixed cell populations on the expansion of each population and on the emergence of neoplastic cells with increase time in vivo will be noted.
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