DEVELOPMENT OF A CELL-LINE FROM SKIN OF GOLDFISH, CARASSIUS-AURATUS, AND EFFECTS OF ASCORBIC-ACID ON COLLAGEN DEPOSITION

Growth characteristics and collagen expression were investigated in GF Sk-S1, a cell line derived from the skin of an adult goldfish (Carassi us auratus). These cells are anchorage dependent, grow well in Leibovi tz-15 medium with 10% fetal bovine serum, and have been subcultured ro utinely for 5 years. Cells at various passages have been successfully cryopreserved and thawed. GFSk-S1 cells show mainly a fibroblastic mor phology at low density, but at confluence islands of epithelial-shaped cells appear among the fibroblastic cells. The cells require little m aintenance, and cultures have been kept viable for more than 3 months without medium changes. Although best growth was observed at room temp erature, cell proliferation still occurred at 28 degrees C, and a subl ine was maintained and passaged fur over a year at 25 degrees C. Cells were exposed to various concentrations ol ascorbic acid, and its effe cts on collagen secretion were monitored by light and electron microsc opy. Under phase-contrast microscopy, confluent CFSk-S1 cells exposed to ascorbic acid at 50 mu g/ml showed distinct development of fibres a s early as 3 days after treatment. Histochemical staining for collagen demonstrated a thick network of fibres under a monolayer of ascorbic acid-treated CFSk-S1 cells, and observation by transmission electron m icroscopy showed collagen fibres with typical banding pattern. This ce ll line appears to show a stable genotype, as collagen expression Mas induced at all passages. GFSk-S1 could be useful for studies not only of regulation of protein synthesis, but also of cell differentiation a nd wound healing.