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  • 1
    ISSN: 1365-2761
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology , Medicine
    Notes: Abstract. Collagenase was used to prepare primary cell cultures from rainbow trout, Oncorhynchus mykiss (Walbaum), gills. Although difficult to subcultivate, one primary culture led to the development of a cell line, RTgill-W1. RTgill-W1 grew in the basal medium, L-15, supplemented with foetal bovine serum at between 5 and 10%, but not in L-15 alone. The cells have been passaged approximately 50 times over a 4-year period. At confluency, the cell shape was predominantly polygonal or epithelial-like. RTgill-Wl cultures were demonstrated by DNA staining with H33258 and by growth on agar to be contaminated with mycoplasma, but this contaminant was eradicated by treatment with mycoplasma removal agent (MRA) and BM cyclin.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1573-6865
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: 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°C, and a subline was maintained and passaged for over a year at 25°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 μg/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
    Type of Medium: Electronic Resource
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