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A marine fish cell line derived from the kidney of red spotted grouper (Epinephelus akaara) named EAGK was established and characterized, this cell line has been subcultured more than 80 passages.The cells multiplied well in Leibovitz's L-15 medium containing 10% fatal bovine serum at temperatures between 25~30℃.The EAGK cell line has a modal diploid chromosome number of 82.The expression of vimentin, desmin and fibronectin can be tested in EAGK cells while cytokeratin can't, with immunofluorescence experiment using four antibodies respectively.