Abstract：Abstract: To compare four methods that were used to isolation and primary culture goat corneal endothelial cells（GCEC）, and research characters of goat corneal endothelial cells in vitro culture. The endothelial cell ,and layers including the Descemet’s membrane were sterilely stripped from 1～2 months guanzhong diary goats cornea, primary culture established by four methods as explant culture，trypsin digestion, dispase digestion and dispase combined trypsin digestion, the ability of dispersion and adhesion of the cells were compared. Results showed that dispase combined trypsin digestion was the optimum method to isolate and purify GCEC.The Morphology, immunostaining, Giemsa staining, NSE staining, scaning and Transmission electron microscope characters proved that the isolated cells were GCEC. It indicated that dispase combined trypsin digestion was optimum method to isolate and purify GCEC, and this method could provide seed cells for theoretical researches of mammalian corneal endothelial cells and tissue engineering corneal endothelium.
