Cultivation of human iris pigment epithelia and identification of its characteristics

Objective:To establish a method to culture a human iris pigment epithelium(IPE) cell line.Methods:Enzyme assisted microdissection was used to isolate IPE cells derived from healthy donor eyes. Separated IPE was then cultured,purified,passaged,freeze preserved and resuscitated. Transmission electron microscope immunochemistry was used to identify and study the characteristics of human IPE.Results:Separated IPE showed as granular or irregular with brown color. Seventy three percent of IPE anchored in 24 h. Anchored IPE spread in multiangular or dental form. After 4～6 circles,the pigment of IPE decreased and even disappeared. Some cells became fibroblast like. Transmission electron microscopy displayed rich microvillus on the surface of IPE with little pigmented grain. Cultured IPE showed as cytokine and S 100 protein positive under immunochemistry study.Conclusion:Enzyme assisted microdissection is a reliable and quick method to establish an IPE cell line.