Production of Sindbis, influenza, and vesicular stomatitis viruses in chicken embryo and rat embryo cell suspensions

Abstract

SummaryStudies were carried out on the production of Sindbis, influenza and vesicular stomatitis viruses in suspensions of chicken embryo and rat embryo cells. The yield of Sindbis virus in chicken embryo cell suspensions was independent of the multiplicity of infection over the range 0.O001 to 0.01 although reduction in mnltiplicity caused a delay in virus production. With influenza virus the use of higher multiplicities gave increased virus yields possibly due to the very slow production at low multiplicities. In both monolayer and suspension cultures of chicken embryo cells addition of serum or use of media richer than minimum essential medium (Eagle) had little effect on Sindbis virus production, but if the glucose were omitted the virus yield was markedly reduced. In cell suspensions, a marked reduction in virus yield occurred if infection were delayed more than 24 hr after cell preparation whereas in monolayers the delay of infection allowed cell propagation and hence a higher yield of virus. It was also shown that vesicular stomatitis virus can be produced in chicken embryo cell suspensions, and that rat embryo primary cell suspensions can be used to prepare both Sindbis and vesicular stomatitis viruses. Sindbis virus obtained from chicken embryo cell suspensions was purified by polyethylene glycol precipitation and sucrose density gradient centrifugation and shown to contain only those proteins previously identified as viral, without any contamination from chicken cell proteins. The relative ease and economics of virus production by cell suspension and monolayer methods is compared.

Autoclavable low cost serum‐free cell culture media. The growth of L Cells and BHK cells on peptones

“…L cells have been used in serum-free media for the production of Venezuelan Equine Encephalitis virus,* and it has been demonstrated that in chick embryo fibroblasts neither sertlm nor amiho acids are necessary for the production of Sindbis virus and that cell growth is not required. 33 It is clear that the nutritional requirements for cell production and virus production may be totally different, and provided that no inhibitors of virus adsorption or replication be present, virus yields may be merely a reflection of cell density irrespective of the medium used.…”