Corneal transparence and thickness mostly depend on corneal endothelial cells. The shortage of transplant-grade donor corneal tissues and limited in vitro expansion of human corneal endothelial cells prompted further impetus for the development of tissue-engineered human corneal endothelium reconstructed in vitro. The culture method of human corneal endothelial cell has been widely used. The standard used to evaluate and identify the human corneal endothelial cells cultivated in vitro has not been established. The objective of this article is to summarize the further study on identification and cultivation of human corneal endothelial cell in vitro.