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

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Abstract
Summary 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 × 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.

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