Serially subcultivated cells as substrates for poliovirus production for vaccine.
Four human diploid cell cultures, WI-38, MRC-5, HEL 299, and IMR-90, and the aneuploid simian cell culture LLC-MK2 were considered in this study as alternatives to primary monkey kidney cells in the production of Inactivated Poliomyelitis Vaccine (IPV). The composition of the basal media used for cell growth and viral replication was found to be critical. Medium CMRL-1969 required a 50% increase in the concentrations of arginine, cystine, glutamine, isoleucine, leucine, methionine, serine, and threonine to produce optimum cell and viral antigen yields, particularly in the Multi-Surface-Cell-Propagator (MSCP) and in the microcarrier suspension culture systems. Also, Medium 199, with Earle's Basic Salt Solution (BSS) for the human diploid cultures, and Hanks' BSS for the simian aneuploid culture, increased the D-antigen yields when compared with other virus growth media deficient in nucleic acid precursors. Approximately 3--4 cell population doublings were obtained in microcarrier culture systems. The average D-antigen yields for poliovirus Types 1,2 and 3 were about 110, 25 and 35, respectively. The physiological requirements for the four human diploid cell cultures were similar indicating easy interchange. The major advantage of the aneuploid LLC-MK2 cells is the reduced serum requirement. The microcarrier culture system is suitable for these cell substrates in the production of IPV provided the microcarriers can be kept in "single-bead" suspension at low rotational speeds of, initially, 10 rpm, with slight increases as the cell density increases.
D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D000956:Antigens, Viral; D002478:Cells, Cultured; D000882:Haplorhini; D006801:Humans; D017955:Poliovirus / Q000254:growth & development*; D014779:Virus Replication
D000596:Amino Acids; D000956:Antigens, Viral