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Growth of High-titre Semliki Forest Virus in Concentrated Suspensions of Chick Embryo Cells

Microbiological Research Establishment, Porton, Salisbury, Wilts.

Freshly prepared chick embryo cells have a lower glycolysis rate than cultured cells and can be used at higher concentrations in suspension to produce virus. Conditions for producing maximal yields (about 10³ p.f.u./cell) of Semliki Forest virus from suspensions containing 10⁷ to 10⁸ cells/ml. were: (1) suitable preparation of the cells, (2) incubation at 31°, (3) maintenance of pH at 7 to 7.4, (4) adequate glucose, (5) adequate oxygen, (6) gentle agitation. The cultures required no nutrient other than glucose and the cells were suspended in Earle's saline. For cell concentrations up to 3 x 10⁷/ml. cultures were stirred in a simple sealed vessel and buffered with NaHCO₃/CO₂. Cultures containing 10⁸ cells/ml. were stirred in a vessel equipped with automatic pH control and produced 10¹¹ p.f.u./ml. of virus.

Received 15 September 1967; accepted 2 October 1967.