Development of a cell line from skin of goldfish, Carassius auratus, and effects of ascorbic acid on collagen deposition.

Abstract

Growth characteristics and collagen expression were investigated in GFSk-S1, a cell line derived from the skin of an adult goldfish (Carassius auratus). These cells are anchorage dependent, grow well in Leibovitz-15 medium with 10% fetal bovine serum, and have been subcultured routinely for 5 years. Cells at various passages have been successfully cryopreserved and thawed. GFSk-S1 cells show mainly a fibroblastic morphology at low density, but at confluence islands of epithelial-shaped cells appear among the fibroblastic cells. The cells require little maintenance, and cultures have been kept viable for more than 3 months without medium changes. Although best growth was observed at room temperature, cell proliferation still occurred at 28 degrees C, and a subline was maintained and passaged for over a year at 25 degrees C. Cells were exposed to various concentrations of ascorbic acid, and its effects on collagen secretion were monitored by light and electron microscopy. Under phase-contrast microscopy, confluent GFSk-S1 cells exposed to ascorbic acid at 50 micrograms/ml showed distinct development of fibres as early as 3 days after treatment. Histochemical staining for collagen demonstrated a thick network of fibres under a monolayer of ascorbic acid-treated GFSk-S1 cells, and observation by transmission electron microscopy showed collagen fibres with typical banding pattern. This cell line appears to show a stable genotype, as collagen expression was induced at all passages. GFSk-S1 could be useful for studies not only of regulation of protein synthesis, but also of cell differentiation and wound healing.