Herein the use of embryonic stem cells of mouse and human origin for studying tumor-induced angiogenesis is demonstrated. Confrontation cultures of embryoid bodies and multicellular tumor spheroids represent the only in vitro model for the investigation of blood vessel and tumor tissue interaction. Previous attempts to study endothelial cell growth into tumor tissue by using multicellular tumor spheroids in confrontation culture with endothelial cells failed due to a lack of endothelial cell invasion (16). In confrontation cultures of embryoid bodies with multicellular tumor spheroids, the time course of endothelial cell invasion into the tumor tissue can be monitored as well as changes in physiological parameters (e.g., intracellular pH, calcium, and reactive oxygen species). The confrontation culture allows changes in protein expression during tumor-induced angiogenesis to be monitored, and may be exploited as in vitro model to study the effects of antiangiogenic agents.

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