A cell line designated SHF-1 was established from fin tissue of the sheepshead, Archosargus probatocephalus. Cells were fibroblastlike and grew best at 26 °C in Leibovitz (L-15) medium containing serum supplements and a final sodium chloride concentration of 0.150 M. The 2N number of primary sheepshead cells was 48 but the SHF-1 line at passage levels above 11 was heteroploid. SHF-1 cells did not replicate any of seven viruses tested. Species confirmation of SHF-1 was accomplished by the cytotoxic antibody test and was attempted with isoelectric focusing of lactic dehydrogenase isoenzymes. Plating efficiency and freeze viability studies were also performed. Key words: fish cell culture, sheepshead, Archosargus probatocephalus, microbiology