A new method for obtaining scanning electron microscopic images of the reorganization process of human dermal microvascular endothelium in vitro

Abstract

A new method for obtaining scanning electron microscopic [SEM] images of the reorganization process of endothelial cells was developed. When covered with a collagen-coated disk, all the cultured endothelial cells reorganized on the collagen of the disk, which was easily taken out from the dish to process for SEM. The reorganization process could be divided into 4 stages: endothelial cell growth (Stage 1), reticular network formation (Stage 2), tubular structure formation (Stage 3), and cytolysis of the tube (Stage 4). Between Stages 1 and 2 the endothelial cells transformed from a cobblestone to a spindle-shaped pattern and fused each other, forming a board-like structure. Between Stages 2 and 3 break up of parts of the board-like structure and outflow of a necrotic mass from the center of the structure occur. At Stage 3 a tubular structure is formed following enwrapping of the cleared center by the surrounding endothelial cells. This method produces a means to study the angiogenesis in a variety of disorders including tumors and wound-healing process using SEM.