The isolation of whole cells from the ciliary epithelium together with some observations on the metabolism of the two cell types

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The cells of bovine ciliary epithelium have been isolated from the underlying stroma and separated into pigmented and non-pigmented fractions. Enzymic digestion of the tissue by papain or pronase, followed by collagenase, was required to free the cells from the tissue and, after filtration, the cell suspension was fractionated by centrifugation on a concentration gradient of colloidal silica. The cells were recovered from the gradient in zones which had only a 10% contamination by the minority cell type. In addition to the enzymic method small sheets of non-pigmented cells were obtained from calf eyes by manual dissection. Respiratory and glycolytic activities were determined and both were found to be higher in the non-pigmented cells than in the pigmented cells. Succinate-tetrazolium reductase activity was also found to be higher in the non-pigmented cells, but lactic dehydrogenase was equally distributed in the two types of cell. The rates of respiration and glycolysis of the pigmented cells were, respectively, little affected by succinate and by anaerobic conditions, whereas the non-pigmented cells showed much greater changes in metabolic activity in response to variations in substrate and oxygen supply. These findings lead to the conclusion that it is the non-pigmented cells that play the major role in the provision of energy for sodium transport by the ciliary epithelium.