In Vitro Angiogenesis Assay for the Guidance of Microvessel Containing Multi-Cellular Lumen Formation

Angiogenesis is a development process of new capillaries from the pre-existing blood microvessel. It involves multiple steps, including proliferation of endothelial cells (ECs), migration, alignment, sprouting, cell-to-cell connection, tube-like structure and lumen formation. One of the major problems faced by the angiogenesis researchers has been the difficulty of finding suitable In Vitro assay to follow the morphological events during angiogenesis development. Morphological events of EC undergoing angiogenesis is of important knowledge for tissue engineering and biomedical societies. In this work we describe step-by-step angiogenesis process and morphological development of human umbilical vein endothelial cells (HUVECs) in the In Vitro three-dimensional fibrin gel assays. Endothelial cell was firstly used to cover polymer fiber surface. When the fiber covered with cells was sandwiched between two fibrin gel containing cells, endothelial cell-covered fiber recruits cells inside the gel to form cell-to-cell connection and tube-like structure. Afterwards cells were degraded their fibrin matrix in order to form complex networks of microvessel containing multi-cellular lumen in between two adjacent fibers.