In vitro cultures of trabecular meshwork cells of the human eye as a model system for the study of cellular aging

Abstract

Cells from the human trabecular meshwork providing a drainage system for the outflow of aqueous humour in the eye were isolated and propagated in monolayer culture. Following serial subcultivation of the primary cultures, there was a gradual decline in the fraction of dividing cells with increasing population doubling level (PDL) resulting finally in growth cessation and disintegration of these ‘senescent’ cultures. The number of population doublings was at most 20. Senescent cultures revealed reduced glycosaminoglycan synthesis rates (as measured by [ 14C]glucosamine incorporation) with a relative decrease of hyaluronic acid and increase of heparan sulfate. Medium-supplied (exogenous) hyaluronic acid enhanced hyaluronic acid synthesis of trabecular meshwork cells cultured in a defined, serum-free medium. Ascorbic acid (25–200 gmg/ml), which is found in very high concentration in the ocular aqueous humour, stimulated hyaluronic acid synthesis of confluent cultures, also. The functional significance of decreased hyaluronic acid (and elevated heparan sulfate) synthesis in the process of cellular aging in vitro (and in vivo?), as well as the importance of hyaluronic acid for the structural integrity and functional activity of the trabecular meshwork were discussed.