[In vitro culture of iris cells

Abstract

The biological behaviour of iris cells in vitro was not yet completely investigated. For a more detailed study of the scope of cultivation of iris cells in vitro we isolated human iris pigment epithelium (IPE) cells and iris fibroblasts. For the purpose of this study iris cells were isolated from 19 donor eyes. A method was established for isolation and cultivation of IPE cells by means of fibronectin coating and the use of a special cell culture medium. Additionally, a method was developed for the selection of fibroblasts from iris stroma (IS) in vitro by means of fibronectin coating, passaging and proliferation in cell culture medium. The IPE and IS cells could be cultivated successfully. The IPE cells started to divide after a mean interval of 5.4 +/- 0.7 days in culture. The mitosis of IS cells was observed after 3.3 +/- 0.87 days in culture. Confluency of IPE cells was reached after 14.7 +/- 4.92 days and by IS cells after 8.1 +/- 1.45 days. Immunocytochemical staining using two antibodies for cytoceratin and one for human fibroblast showed that the IPE cell culture was pure and that the IS culture consisted of fibroblasts. Furthermore, electron microscopy of IPE and IS cultures confirmed the results of the immunocytochemical staining. The use of human IS and IPE cells in vitro has established a novel model for the research on iris cells. The model might possibly be applied in the research of metabolic structures and diseases of the iris.