Hepatitis B-virucidal testing of
biocides in quantitative
suspension tests using duck hepatitis B virus (DHBV) requires primary duck embryonic hepatocytes for viral propagation. To improve the test system and availability of these cells, commercial culture plates with different growth surfaces were tested for cell cultivation and different approaches for cryopreservation of hepatocyte
suspension were examined. After 12 days of culture, the largest amounts of hepatocytes were grown in CellBIND and
TTP plates and CellBIND surface showed the lowest tendency of monolayer detachment nearly comparable with
collagen 1-coated CELLCOAT plates. For cryopreservation of hepatocyte
suspension, the use of growth medium supplemented with
fetal calf serum (FCS) and
dimethyl sulfoxide (ME(2)SO), FCS supplemented with ME(2)SO or cryosafe-1 as
cryoprotective agents provided the highest rates of surviving cells after thawing. The freezing-thawing process did not significantly reduce the susceptibility of hepatocytes to
infection with DHBV. In conclusion, plates without
collagen 1 such as CellBIND are recommended for cultivation of primary duck embryonic hepatocytes in infectivity experiments of DHBV for virucidal testing of
biocides. The use of cryopreserved hepatocytes is possible when freshly isolated cells from the liver of duck embryos are not available.