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Blood serum model and cell model of in vitro HBV (Hepatitis B Virus) infection and building method thereof

A serum model and in vitro infection technology, applied in the field of animal cell lines, can solve the problems of difficult establishment of infection, poor interpretation of HBV infection, and unclear research on hepatitis B virus, etc., and achieve the effect of simple method and wide source

Inactive Publication Date: 2013-08-14
TONGJI HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, since the establishment of the cell line was completed with the assistance of DMSO and dexamethasone, the mechanism of HBV infection cannot be well explained
[0006] Although HBV is highly infectious in vivo, it is difficult to establish effective infection in cell culture in vitro
The research on the early stage of hepatitis B virus, such as the recognition of HBV and molecules adsorbed to liver cells, etc. is still not very clear. Most of the existing data on the early infection of hepatitis B virus come from the research on DHBV

Method used

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  • Blood serum model and cell model of in vitro HBV (Hepatitis B Virus) infection and building method thereof
  • Blood serum model and cell model of in vitro HBV (Hepatitis B Virus) infection and building method thereof
  • Blood serum model and cell model of in vitro HBV (Hepatitis B Virus) infection and building method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1: Establishment of serum model of hepatitis B virus infection in vitro:

[0053] 1. Cell culture:

[0054] 1. Cell recovery: Take out HepG2 and Hep2.2.15 cells from the liquid nitrogen tank, quickly place them in a constant temperature water bath at 37°C, and shake them constantly. After the cryopreservation solution is dissolved, wipe the surface of the cryovial with 75% alcohol. Add 10mL DMEM medium to the centrifuge tube, centrifuge at 1000rpm for 10 minutes, discard the supernatant in the centrifuge tube in the ultra-clean table, add 10mL DMEM medium to the cell culture flask, and HepG2.2.15 cell culture medium needs to be added G418 200ug / mL. Put at 37℃, 5%CO 2 In the cell culture incubator, change the medium or passage depending on the growth of the cells.

[0055] 2. Passaging of cells: The old HepG2.2.15 cell culture medium was collected based on a 15mL centrifuge tube, and the old HepG2 cell culture medium was discarded. Wash 3 times with sterile P...

Embodiment 2

[0069] Example 2 Verification of the cell model of hepatitis B virus infection in vitro

[0070] The serum model formed by mixing the deviralized hepatitis B big three positive serum and HepG2.2.15 virus particles was used as the experimental group, the normal human serum and the virus mixture of HepG2.2.15 cells were used as the negative control group, and the HepG2 cells treated with DMSO for six days were used for the experiment. The serum of the group was used as the positive control group. See below for details figure 1 :

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Abstract

The invention discloses a blood serum model and a cell model of in vitro HBV (Hepatitis B Virus) infection and a building method thereof. The blood serum model is formed by mixing virus-removed hepatitis B great three positive blood serum with HepG2.2.15 viral particles, wherein the copy number of the virus-removed hepatitis B great three positive blood serum is that HBV-DNA (deoxyribonucleic acid) >10<8>copies / mL. The cell model is formed by co-culturing normal hepatic cell / hepatoma cell and the blood serum model. The models disclosed by the invention have the advantages that the cell is wide in source and easily available; the general characteristic of the hepatoma cell is maintained; and the hepatoma cell can be limitlessly subcultured. The method disclosed by the invention is simple and convenient and capable of well simulating the natural process of HBV entering the cell.

Description

technical field [0001] The invention belongs to the field of animal cell lines, in particular to a serum model and a cell model of hepatitis B virus infection in vitro and a method for establishing the same. Background technique [0002] There are hepatitis B virus (HBV) infected people all over the world. In Asia, Africa, the Pacific and other regions, hepatitis B virus HBV is one of the main pathogens that endanger human health and cause human liver disease. China is a high incidence area of ​​hepatitis B. The persistent infection of liver cells by hepatitis B virus leads to abnormal liver function in some people and eventually develops into relatively serious liver diseases, including liver cirrhosis and hepatocellular carcinoma. The current anti-HBV drugs such as interferon and nucleoside analogs limit their wide clinical application due to serious side effects. Studying the life history of HBV and its pathogenic mechanism, especially the interaction between the host an...

Claims

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Application Information

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IPC IPC(8): C12N5/071C12N7/00
Inventor 廖家智孙淑珍林菊生何星星黎培员
Owner TONGJI HOSPITAL ATTACHED TO TONGJI MEDICAL COLLEGE HUAZHONG SCI TECH
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