A male, full-term Atlantic bottlenose dolphin (Tursiops truncates) calf was delivered stillborn, apparently as a result of complications during the delivery process. Tissue specimens were obtained from the skin, lung, kidney, and deep muscle within approximately 30 minutes after delivery. A portion of each type of tissue was diced and used to prepare explant cultures, and the remaining portions of the samples were digested with trypsin (an exception was the skin specimen which was only used for explant cultures). Samples of the trypsinized tissues were removed after various lengths of digestion, washed and placed in one of two growth media and plated out in plastic tissue culture flasks. Basal media employed included a carbonate buffered medium (MEM) and a phosphate buffered medium (L-15). Complete growth medium consisted of basal medium supplemented with 5-20% fetal bovine serum, non-essential amino acids, vitamins, l-glutamine, and an antibiotic/antimycotic mixture. In some cases fibroblastic growth factor or epithelial growth factor was added. No growth was ever observed with lung or skin cultures in MEM, and no growth of muscle cells was observed using either MEM or L-15. Although cell growth in some kidney cultures (e.g. five hour trypsin digests) in both MEM and L-15 was observed in the first two weeks after initiation of cultures, the cultures did not thrive in MEM, and eventually these cultures were switched to L-15. In kidney cultures (five hour trypsin digest only) in L-15, growth sufficient to begin subculture and expansion of the culture was achieved. In lung explants cell growth was first observed at day 30 and in the skin explant at day 40 (in both cases in cultures with L-15, as indicated). Growth of lung and skin cultures in L-15 sufficient to begin subculture and expansion of the cultures was achieved (although growth rate and general condition of cells in skin cultures were poor). Aliquots of the lung, kidney, and skin cultures (now primary cell lines) were frozen in liquid nitrogen for storage, and efforts were commenced to develop permanent cell lines from the primary lines by a series of subcultures. The lung cell line is presently in passage 32 and the kidney cell line, which was only recently removed from frozen storage for further subculture, is presently in passage 9. The skin primary cell line is being stored frozen at passage 5 and below pending future development. Both the kidney and lung lines have a generally fibroblastic appearance. Work is currently underway to characterize the lines with respect to population doubling time, plating efficiency, karyotyping, susceptibility to viral pathogens, etc. The lines have potential for use as viral screening/diagnostic hosts for cetacean viruses, or for studies of viral replication in cetacean cell systems.