Viral antigen production in cell cultures on microcarriers Bovine parainfluenza 3 virus and MDBK cells.

摘要: Viral antigens can be obtained from infected mammalian cells cultivated on microcarriers. We have worked out parameters for the production of bovine parainfluenza 3 (PI-3) virus by Mandin-Darby Bovine Kidney (MDBK) Cytodex 1 microcarriers (MCs) in spinners flasks and bioreactor using fetal serum (FBS) supplemented Eagle minimal essential medium (Eagle-MEM). Medium renewal during cell culture was shown to crucial optimal MCs loading (>90% with confluent monolayers) growth (2.5 x 10(6)cells/mL a micro(x) (h(-1)) 0.05). Since cultures performed lower amount (1g/L), showed good performances terms loading, we designed batch experiments concentration view optimizing production. Studies concentrations (0.85 g/L) that an increase initial seeding (from 7 40 cells/MC) led different kinetic but comparable final ((8-10)x10(5)cells/mL at 120 h) (210-270 cells/MC). Upon infection PI-3 virus, decrease MC directly related multiplicity (moi) used infection. Infected also higher consumption glucose lactate. The antigen among not significantly attained values ranging from, respectively, 7-9 log(10) TCID(50)/mL 1.5-2.2 OD. kinetics sharp first 24h those increased after 24h. differential explained sensitivity temperature. In establishing protocol based previous experiments, MDBK under perfusion 1.2L were 12L 12 TCID(50). Other than 1, are proposed as basis approaching development bioreactors.