Quantitative analysis of unidirectional 2-D tissue formation of endothelial cells

We developed a reliable and quantitative method for measuring the dynamic process of unidirectional two-dimensional (2-D) tissue formation of endothelial cells (ECs) in vitro. The culturing of bovine ECs in an assembled culture chamber provided a square monolayered cell sheet with a linear margin when disassembled at the confluency. The cell sheet maintained in culture showed a unidirectional which was determined from the daily observation of tissue, allowed us to determine quantitatively the dynamic process of unidirectional endothelialization in vitro. The endothelialized distance and the endothelializing zone on a glass slide were found to be nearly 500 microns/day and 750 microns in width, respectively. Thus, the method developed here provided information of the 2-D tissue formation process. This model would be useful as an in vitro model which simulates the anastomotic endothelialization of an artificial vascular graft.