Culture of Urethral Fibroblasts: Cell Morphology, Proliferation and Extracellular Matrix Synthesis

The in vitro morphology and proliferative activity of urethral fibroblasts were characterized from primary isolates of fetal bovine urethral spongiosum. Cells were identified as fibroblastic based on their spindle-shaped morphology and lack of expression of phenotypic markers characteristic of either urethral smooth muscle or epithelial cells. Urethral fibroblasts proliferated at a steady rate and did not exhibit contact-inhibition of growth in a culture medium of Medium 199 supplemented with 10% newborn calf serum. Both immunohistochemistry and metabolic labeling analyses of extracellular matrix synthesis demonstrated the presence of collagen types I and III, and fibronectin. Quantitation of the collagenous components of the cell medium revealed that urethral fibroblasts synthesize 85% type I collagen, 14% type III collagen and < 1% type V collagen. In contrast, the urethral fibroblast cell layer matrix consisted of 98% type I collagen, < 1% type III and < 1% type V collagen.