Arbovirus Replication in Mosquito Cell Lines (Singh) Grown in Monolayer or Suspension Culture

Summary Singh's mosquito cell line derived from larval A. albopictus was grown in spinner cultures with a doubling time of 24-30 hr at 23 or 28°. It was found to be adaptable to various partially defined growth media. The typical chromosome count was 6, with 12 chromosomes appearing in 25% of figures. Infection of the cells with Sindbis virus or type 2 dengue virus at multiplicities of 1 or 2 pfu/cell and maintenance at 28° yielded viral growth cycles similar in timing to those seen in infected vertebrate cells at 37°. Cells chronically infected with Sindbis virus yielded virus at the rate of 1 pfu/cell/-day 7 weeks after infection. Sindbis virus production by cells exposed to actinomycin-D was unaffected by noncytotoxic doses. CPE due to actinomycin occurred at concentrations similar to those affecting vertebrate cells.