| Hepatitis B is a chronic illness caused by viruses, and it can not be healed completely. What's more, hepatitis B can deteriorate to cirrhosis and hepatocellular carcinoma with the development of illness. There are many people infected with hepatitis B virus (HBV) in China. So it is of great importance that studies on HBV is carried on. From Blumbeg having found HBV surface antigen in 1965 to having completed clone and sequence of HBV DNA genome in 1997, Researchers have been making great progress in makeup of HBV genome, structure and biological traits of antigens, and life circle of HBV. Due to deficiency of multiplying system and valiable animal and cell models, little was known about the early mechanism of initiation of infection. Consequently, basic and clinical research on hepatitis B and other hepatitis concerned were effected some way. Among the means developed recently, cell model was one of ideal ways to make sure the infection mechanism. Because of the limitness of human hepatic tissue and strong preference of HBV to hosts, Tupaia was choosed to provide primary hepatocytes, which have the same infection activities as those of human hepatocytes. There were two important parts in experiments: one was isolation and primary culture of hepatocytes, the other was to determine the best way to infect hepatocytes efficiently. Primary hepatocytes were prepared by two-step collagenase perfusion. Then cells were cultured in DMEM medium with 10% fetal bovine serum, with the result of high plating efficiency and excellent function maintenance. Finally it was determined that when serum with HBV was added in dilution of 1:50-1:300, cells of density of 10s per mililitre could be infected sufficiently in cell medium with 2% DMSO and 4% PEG two days after cell plating. The cell model was useful for evaluating potential genetic engineering vaccine and for studying on HBV DNA replication. |
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