[In vitro infection of human hepatoma (Hep G2) cell line by hepatitis B virus positive serum].
OBJECTIVE
To establish a culture system of HBV positive serum infected Hep G2 cells in vitro.


METHODS
Hep G2 cells were seeded into six-well cluster dishes, at 1 x 10(-6) cells per well and incubated with 3 ml 10% fetal calf serum/ Dulbecco's modified Eagle's medium (10% FCS/DMEM) at 37 degrees in 5% CO2 air. At 24 h after plating, infection group Hep G2 cells were cultured with 0.5 ml HBV positive serum, in control group HBV negative serum was used, 24 h later the inoculums was removed. The cells were then extensively washed with 0.01 mol/L phosphate-buffered saline (PBS). After washing with PBS, 4 ml 2% FCS/DMEM were added to each well and the medium was collected every 12 h. ELISA method was used to detect HBsAg in culture medium. HBV DNA in cells and culture medium was detected by PCR.


RESULTS
In infection group, HBsAg could be detected from cell culture medium from 12 h (after PBS washed) to 84 h. HBV DNA could be detected by PCR in culture medium and cells.


CONCLUSIONS
Infection of Hep G2 cells by HBV positive serum is feasible.
Wang
An-hui
AH
Department of Epidemiology, School of Preventive Medicine, Fourth Military Medical University, Xi'an 710033, China.
Men
Ke
K
Xu
De-zhong
DZ
Yan
Yong-ping
YP
Lu
Juan
J
Zhang
Jing-xia
JX
D006528:Carcinoma, Hepatocellular / Q000473:pathology / Q000821:virology; D045744:Cell Line, Tumor; D004279:DNA, Viral / Q000235:genetics; D004797:Enzyme-Linked Immunosorbent Assay; D006509:Hepatitis B / Q000097:blood / Q000821:virology*; D006514:Hepatitis B Surface Antigens / Q000032:analysis; D006515:Hepatitis B virus / Q000235:genetics / Q000254:growth & development* / Q000276:immunology; D006801:Humans; D008113:Liver Neoplasms / Q000473:pathology / Q000821:virology; D016133:Polymerase Chain Reaction; D044967:Serum / Q000821:virology
D004279:DNA, Viral; D006514:Hepatitis B Surface Antigens