Hepatitis B virus infection and replication in primarily cultured human fetal hepatocytes.
OBJECTIVE
To investigate the infection and replication of hepatitis B virus (HBV) in primarily cultured human fetal hepatocytes (HFHs).


METHODS
The human fetal hepatocytes were cultured in serum-free medium, HBV-positive serum was added into the medium to study the susceptibility of hepatocytes to HBV infection. The supernatant was collected for ELISA assay of HBsAg and HBeAg, and quantitative fluorescence PCR for HBV-DNA assay daily. Albumin and HBcAg, CK8 and CK18 expressions were detected by immunohistochemistry in cultured hepatocytes. Content of lactate dehydrogenate (LDH) was measured to find out the integrity of the cell membrane.


RESULTS
A stable hepatocyte culture system was established. HBV could infect the hepatocytes and replicate, and HBcAg expression could be detected by immunohistochemistry in hepatocyte-like cells. HBV-DNA in the supernatant could be detected from d 2 to d 18 and HBsAg and HBeAg were positive on d 3-d 18 after HBV infection. HBV in medium increased from d 0 to d 6 and subsequently decreased as the cells were progressively loosing their hepatocyte phenotypes.


CONCLUSIONS
HBV could infect human fetal hepatocytes and replicate. This in vitro model allowed a detailed study on early events associated with human HBV entry into cells and subsequent replication.
Lin
Min
M
Chaozhou Central Hospital, Chaozhou 521021, Guangdong Province, China.
Chen
Qun
Q
Yang
Li-Ye
LY
Li
Wen-Yu
WY
Cao
Xi-Biao
XB
Wu
Jiao-Ren
JR
Peng
You-Peng
YP
Chen
Mo-Rui
MR
D000956:Antigens, Viral / Q000378:metabolism; D002478:Cells, Cultured; D004279:DNA, Viral / Q000235:genetics / Q000378:metabolism; D006509:Hepatitis B / Q000378:metabolism / Q000473:pathology*; D006512:Hepatitis B Core Antigens / Q000378:metabolism; D006513:Hepatitis B e Antigens / Q000378:metabolism; D006515:Hepatitis B virus / Q000235:genetics / Q000276:immunology / Q000472:pathogenicity*; D022781:Hepatocytes / Q000276:immunology / Q000378:metabolism / Q000821:virology*; D006801:Humans; D008099:Liver / Q000166:cytology / Q000196:embryology*; D014779:Virus Replication / Q000502:physiology*
D000956:Antigens, Viral; D004279:DNA, Viral; D006512:Hepatitis B Core Antigens; D006513:Hepatitis B e Antigens