Primary cell cultures (n = 16) were initiated from tissues of embryonic and neonatal larval Ornithodoros moubata following methods developed for hard ticks. After maintenance for 20-25 months in vitro, cell multiplication commenced in surviving cultures, leading to the establishment of six cell lines designated OME/CTVM21, 22, 24, 25, 26 and 27. All lines are maintained at 28 degrees C, with subculture at 2-8 week intervals. The cultures comprise heterogeneous populations of large cells of 15-100 microm in diameter, often with finger-like protrusions and/or intracellular crystals, rarely attached, predominantly floating and forming clumps or hollow multicellular vesicles up to 1 mm in diameter. Attempts to cryopreserve the cells are described. Tick-borne encephalitis virus has been serially passaged ten times in OME/CTVM21 cells without significant decrease in virus production and with no change in its biological properties as shown by the size and morphology of plaques produced in porcine kidney cells.