Controlling physiological angiogenesis by hypoxia-induced signaling

Abstract

The full sequence of signals leading to new blood vessel formation is a physiological response to tissue hypoxia through upregulation of angiogenic factor cascades. Controlled initiation of this mechanism for therapeutic/engineered angiogenesis must rely on precisely localized hypoxia. Here we have designed a 3D in vitro model able to test the effect and predictability of spatially positioned local hypoxic stimuli using defined cell depots within a 3D collagen matrix. Cell-mediated hypoxia was engineered using human dermal fibroblasts (HDFs), to generate a local population of Hypoxia-Induced Signaling ( HIS) cells. HIS cell depots released angiogenic factors which induced directional endothelial cell (EC) migration and tubule formation in a spatially defined assay system. Non-hypoxic baseline control cultures induced minimal EC migration with little tubule formation. Furthermore, depots of HIS cells, positioned in the core of 3D collagen constructs directed host vessel in-growth deep into the implant by 1 week, which was at least 7 days earlier than in non-hypoxia pre-conditioned constructs. The functionality of in vivo vascularisation was verified by real-time monitoring of O 2 levels in the core of implanted constructs. These findings establish the angiogenic potential of HIS cells applicable to in vitro tissue modeling, implant vascularization and engineering predictable angiogenic therapies.