A microcarrier cell culture system for large scale production of hepatitis A virus.
Hepatitis A virus (HAV) was isolated from human faeces using a fetal rhesus monkey kidney cell line (Frhk-4). Infectious medium from passage 12 was used to inoculate a large (5000 cm2) microcarrier cell culture maintained in suspension. The microcarriers used were swollen, collagen-coated dextran beads on which it was easy to propagate Frhk-4 cells. Intra- and extra-cellular virus levels were assayed and compared with conventional cultures in 25 cm2 plastic flasks. The results show that virus production per cell was similar in both systems. The number of cells per area unit in confluent cultures was initially lower in the microcarrier culture but subsequently increased. Two to three weeks post inoculation the virus yield per area unit in the microcarrier system was half of that of the conventional culture. The lower cell density per area unit in the microcarrier system was compensated by the large growth area that could be maintained in a single vessel and the total production of virus was substantial. Weekly harvests of medium with HAV antigen titres around 10(-2) contained antigenic material sufficient for several thousands of anti-HAV IgM tests. Propagation of HAV in microcarrier cell cultures thus seems a safe and simple way to produce large amounts of HAV.
D000818:Animals; D002452:Cell Count; D002455:Cell Division; D002460:Cell Line; D003094:Collagen; D003470:Culture Media; D003911:Dextrans; D006507:Hepatovirus / Q000254:growth & development*; D008253:Macaca mulatta; D008863:Microspheres; D014776:Virus Cultivation / Q000379:methods*
D003470:Culture Media; D003911:Dextrans; D003094:Collagen