Study of Viral Vectors in a Three-dimensional Liver Model Repopulated with the Human Hepatocellular Carcinoma Cell Line HepG2

作者： Thomas Hiller1, Viola Röhrs1, Eva-Maria Dehne2, Anke Wagner1,3, Henry Fechner1, Roland Lauster2, Jens Kurreck1 1Department of Applied Biochemistry,Institute of Biotechnology, Berlin University of Technology, 2Department of Medical Biotechnology,Institute of Biotechnology, Berlin University of Technology, 3Department of Bioprocess Engineering,Institute of Biotechnology, Berlin University of Technology

简介：

Overview

This study presents a method for developing a three-dimensional liver model using a recellularized extracellular matrix from decellularized rat livers. This model allows for the investigation of viral vectors and gene transfer in a humanized environment, enhancing the relevance of findings to human physiology.

Key Study Components

Area of Science

• Neuroscience
• Biology
• Viral vector research

Background

• The use of decellularized organs as scaffolds for human cell studies.
• Importance of studying biological processes in a human-like environment.
• Reduction of animal testing through advanced ex vivo models.
• Potential applications in gene therapy and cancer research.

Purpose of Study

• To create a humanized liver model for studying viral vectors.
• To improve animal welfare by minimizing the use of live animals.
• To facilitate research on gene transfer and infectious viruses.

Methods Used

• Expansion of Hep G2 hepatic cell line for recellularization.
• Use of a bioreactor system for perfusion and cell seeding.
• Monitoring of physiological parameters during culture.
• Transduction of the liver model with AAV vectors for gene expression analysis.

Main Results

• Successful recellularization of the liver ECM with Hep G2 cells.
• Quantification of vector genomes indicating effective gene transfer.
• Demonstration of the model's utility for studying viral vectors.
• Validation of gene expression through RT-PCR and western blotting.

Conclusions

• The recellularized liver model is a promising tool for virology research.
• This method supports ethical research practices by reducing animal use.
• Further applications in gene therapy and cancer research are anticipated.

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