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Table 1. Summary of HBV in vitro hepatocyte culture models

From: Advances in HBV infection and replication systems in vitro

Classification Cell line Advantages Shortcomings HBV infection rate and
application of the models
HBV replication cell lines (1) HepG2.2.15 cells cccDNA accumulation
Stable and continuous HBV gene expression and replication
Low viral replication level
Antigen expression instability
Virions are produced from the integrated DNA
Screening and evaluation of antiviral drugs, etc. [90].
  (2) HepAD38 (EF9,EFS19) cells Cells differentiate quickly
Produce high titers of viral particles
cccDNA accumulation
Hepatoma cells stably expressing HBV from a Tet-on/Tet-off system
Incomplete viral life cycle
Virions are produced from the integrated DNA
Screening and evaluation of antiviral drugs, etc.
A potential source for tissue culture derived virions [91].
  (3) Ad-HBV1.3-systems No species barrier
Efficient expression of HBV
HBV expression and mutation can be controlled
Direct observation of transfection and infection efficiency (integrated green fluorescent protein gene)
Missing HBV natural infection stage Used to establish animal models of acute hepatitis B infection [92].
  (4) HBV baculovirus system Easy detection of riboprotein-bound HBV DNA
High HBV replication level
Formation of infectious viruses and a detectable intracellular cccDNA pool
Nonreceptor-mediated entry
Gene transfer is restricted to certain species
Missing HBV natural infection stage
Quantify the effect of antiviral agents on nuclear HBV DNA
Used for studying the resistance of HBV to nucleoside analogs [93].
Cell lines that can be infected with HBV (1) Human fetal hepatocytes Phenotypically and biologically functionally close to primary adult human hepatocytes Low infection efficiency
Short infection time
Limited availability
Large donor-donor variations
HBV infection rate12%-90% [22, 94].
Coculturing with hepatic non-parenchymal cells and subsequent addition of 2% DMSO leads to the formation of hepatocyte islands with prolonged phenotypic maintenance [25].
The early events in viral entry into cells as well as viral replication [23].
  (2) Adult human hepatocytes The gold standard host cell to HBV infection experiments
Closest to the physiological characteristics of hepatocytes in vivo
Close to the natural process of infection
Limited life cycle
Unpassable culture
Phenotypically unstable in vitro
Rapidly lose permissiveness for HBV infection
Large donor-donor variations
HBV infection rate 20%-100% [26, 28].
Used for studying the process of HBV infection [5, 28].
Studying on apoptosis [26].
Preparation of 3D primary hepatocyte culture system for analyses of liver diseases, drug metabolism, and toxicity [40, 41].
  (3) Co-culture system Test the utility of various direct-acting antivirals (DAAs) and putative host-targeting antivirals (HTAs);
Assessing preclinically the efficacy of other entry inhibitors and possibly (vaccine-induced) neutralizing antibodies;
Wide variability between donors in terms of HBV permissiveness Inflammation and drug-Induced Hepatotoxicity [95].
  (4) Primary Tupaia hepatocytes The only species susceptible for HBV infection besides humans and chimpanzees Expensive HBV infection rate >70% [52].
Used for in vitro as well asin vivo infection experiments [96].
HBV specific receptor identification [78].
  (5) HepaRG cells Preserve the specific functional properties of hepatocytes
Support the complete HBV life cycle
Produce HBV cccDNA
Strict culture conditions
Low infection efficiency
HBV infection rate <30% [56, 78].
HBV molecular mechanism and screening, evaluation of anti-HBV drugs; cccDNA spread etc. [57].
Drug metabolism and toxicity [58, 59].
  (6) In vitro systems based on induced pluripotent stem (iPS) cell-derived human hepatocytes Biological characteristics similar to those of normal liver cells
Support the complete life cycle of the virus
Complete natural immune system
Complicated operation HBV infection rate 25% [97].
Drug hepatotoxicity screening [98].
The life cycle of HBV virus and virus-induced hepatic dysfunction [66].
  (7) NTCP overexpressing hepatoma cell lines Support the complete life cycle of the virus
Flexibility and easy handling
Low susceptibility to serum-derived HBV
The multiplicity of infection (MOI) needed to achieve infection is extremely high
No substantial viral spreading following infection
HBV infection rate 50% [99].
Large-scale screening of antiviral drugs for targeting NTCP [91].