True lymphovascular invasion in most carcinomas adversely affects survival, but in vitro human models of lymphatic invasion are lacking. Herein is discussed one such model in which certain mechanistic aspects of lymphatic dissemination can be investigated easily in any laboratory. S100P expression is known to be significantly upregulated in the lymph node metastases of human pancreatic adenocarcinoma. Thus, an S100P overexpressing cell line (S5L) and a vector control (V3L) pancreatic cancer cell lines were used to develop a co-culture model with human dermal lymphatic endothelial cells. Adhesion, the effect of cancer cells on lymphatic permeability and trans-lymphatic endothelial migration could be reproducibly quantified in this model and parametric statistical analysis confirmed significant differences between the two cancer cell lines in these functional assays. This proof-of-principle study demonstrates that this simple model can be used to assess lymphatic invasion in a human cancer context in real-time in vitro.