Rabies virus production in non-woven polyester fabric(NWPF) packed-bed reactors

The production of rabies virus from baby hamster kidney-Ankara66 (BHK-An(66), HUKUK 99050302) monolayer cells was examinedin a packed-bed reactor containing non-woven polyester fabric (NWPF)discs. At first, growth characteristics of the cells were determined instatic culture. The suspension culture studies were realized in a 1litre spinner basket with NWPF support. BHK-An(66) cells(inoculation density, 5x10(4) cells.ml(-1)) were maintained in the reactor loaded with5 g.l(-1) carrier. The culture medium wasEagle's minimal essential medium supplemented with 10% (v/v)fetal bovine serum at 37 degrees C. During the culture, the mediumwas sampled daily to assess glucose and lactate concentrations. At theend of the 7 day culture period the cell density was found to be2.2x10(7) cells.ml(-1), and thereactor was inoculated with 5 ml [1.7x10(6) focus-forming units (ffu).ml(-1)] of the CVS 11(Challenge Virus Standard) strain of rabies virus. After a 72 hincubation period, the cultures were stained with fluorescein-conjugated anti-rabies globulin and were observed using a fluorescencemicroscope. Virus titres determined by the Spearman-Kärber method were 2.2x10(5) ffu.ml(-1). In conclusion, NWPF packedreactors can be considered as a suitable system for the large-scaleproduction of rabies virus.