The synthesis of glycosaminoglycans by cultures of rabbit corneal endothelial and stromal cells

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Abstract
Confluent monolayer cultures of rabbit corneal endothelial and stromal cells were incubated independently with [35S]sulfate and [3H]glucosamine for 3 days. After incubation, labeled glycosaminoglycans were isolated from the growth medium and from a cellular fraction. These glycosaminoglycans were further characterized by DEAE-cellulose column chromatography and by sequential treatment with various glycosaminoglycan-degrading enzymes. Both endothelial and stromal cultures synthesized hyaluronic acid as the principal product. The cell fraction from the stromal cultures had significantly less hyaluronic acid than that from the endothelial cultures. Both types of cells synthesized a variety of sulfated glycosaminoglycans. The relative amounts of each sulfated glycosaminoglycan in the 2 cell lines were similar, with chondroitin 4-sulfate, chondroitin 6-sulfate and dermatan sulfate as the major components. Heparan sulfate was present in smaller amounts. Keratan sulfate was also identified, but only in very small amounts (1-3%). The presence of dermatan sulfate and the high content of hyaluronic acid are similar to the pattern of glycosaminoglycans seen in regenerating or developing tissues, including cornea.