title: Comparison of initial feasibility of host cell lines for viral vaccine production.
abstract: In order to reduce the time required for the development and production of viral vaccines, host cell lines should be available as expression systems for production of viral vaccines against groups of viral pathogens. A selection of cell lines was compared for their initial feasibility as expression system for the replication of polioviruses, influenza A viruses and respiratory syncytial virus (wild type strain A2). Six adherent cell lines (Vero, HEK-293, MRC-5, CHO-K1, BHK-21 c13, MDCK) and six single cell suspension cell lines (CAP, AGE1.CR.HS, sCHO-K1, BHK-21 c13 2p, MDCK SFS) were studied for their ability to propagate viruses. First, maximum cell densities were determined. Second, virus receptor expression and polarization of the cell lines regarding receptor distribution of eight different viruses were monitored using flow cytometry and immunocytochemistry. Organization of the actin cytoskeleton was studied by transfection of the cells with Lifeact™, a construct coding for actin-EGFP. Finally, the ability to produce virus progeny of the viruses studied was assayed for each cell line. The results suggest that single cell suspension cell lines grown on serum free medium are the best candidates to serve as host cell lines for virus replication.
authors.affiliation: Institute for Translational Vaccinology, PO Box 450, 3720 AL, Bilthoven, The Netherlands. Danielle.Vlecken@intravacc.nl
authors.forename: Danielle H W
authors.initials: DH
authors.lastname: Vlecken
authors.forename: Ralf P M
authors.initials: RP
authors.lastname: Pelgrim
authors.forename: Slawomir
authors.initials: S
authors.lastname: Ruminski
authors.forename: Wilfried A M
authors.initials: WA
authors.lastname: Bakker
authors.forename: Leo A
authors.initials: LA
authors.lastname: van der Pol
mesh_terms: D000818:Animals; D002460:Cell Line; D049109:Cell Proliferation; D003599:Cytoskeleton / Q000378:metabolism; D005434:Flow Cytometry; D006801:Humans; D007150:Immunohistochemistry; D009980:Influenza A virus / Q000254:growth & development; D017955:Poliovirus / Q000254:growth & development; D011991:Receptors, Virus / Q000032:analysis; D012136:Respiratory Syncytial Viruses / Q000254:growth & development; D013678:Technology, Pharmaceutical / Q000379:methods*; D014765:Viral Vaccines / Q000302:isolation & purification*; D014776:Virus Cultivation / Q000379:methods; D014779:Virus Replication
chemical_list: D011991:Receptors, Virus; D014765:Viral Vaccines
keywords: Host cell line; Influenza; Polio; RSV; Virus cultivation; Virus receptor