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Isogeneic ginbuna crucian carp is a naturally occurring gynogenetic fish. Some of the advantages to using this clonal fish are that they are easily maintained, they are fertile, and they are sufficiently similar to mice in their yearling size (40-60g ) to use in immunological experiments, especially for syngeneic studies, in comparison with other artificial gynogenetic and/or inbred fishes. In addition, several clones that are allogeneic to one another have been established. These clonal fishes have been used to investigate adoptive transfer of immunity1),2) and graft rejection3). For the study of T cell mediated immunity, inbred animals and/or cell lines derived from them are essential because antigen recognition by T lymphocytes is genetically restricted by the major histocompatibility complex (MHC). In particular, their cell lines are useful as target cells to detect T cell-mediated cytotoxicity. In fish, however, to our knowledge, neither clonal fish nor their cell lines have been simultaneously established. We tried to establish a cell line from inbred fish and have succeeded in passaging and maintaining a cell line of isogeneic ginbuna crucian carp Carassius auratus langsdorfii through the 92nd passage. Female and yearling isogeneic ginbuna crucian carp (approximately 45g), Carassius auratus langsdorfii, which originated from Lake Kasumigaura (K1 clone), were obtained from the National Research Institute to Aquaculture. Caudal fins were cut from the fish, bathed in 1% sodium hypochlorite solution for 1 min, and then rinsed in 70% ethyl alcohol for a few seconds. The fins were cut into approximately 1-mm cubes with sterile scissors, and the cubes were transferred to 25cm<SUP>2<SUP> culture flasks (Corning Labs. Inc.), and allowed to dry for 2h so that they would stick to the inside surface of the flasks. Culture medium (MEM-10) was prepared from Eagle's minimum essential medium (MEM; Nissui Pharmaceutical Co.) supplemented with 10% fetal bovine serum (FBS; Commonwealth Serum Laboratories), 4.5mM sodium bicarbonate (NaHCO3), 4mM L-glutamine, and 14 mM N-[2 Hydroxyethyl]piperazine-N'-[2-ethanesulfonic acid] (HEPES; Sigma Chemical Co.). Flasks containing the fin cubes were incubated at 25•Ž with MEM-10 and the cells were subcultured at confluency using a dispersant of 0.1% trypsin (DIFCO Laboratories) in phosphate-buffered saline (PBS) containing 0.02% ethylenediaminetetraacetic acid disodium salt (EDTA). These cells were designated CFK (Carassius fin from Lake Kasumigaura) cells. To determine the cell growth, MTT assays described by Denizot and Lang4) were used with slight modifications. MTT

A Cell Line (CFK) from Fin of Isogeneic Ginbuna Crusian Carp.