Functionality of the in vitro hepatic models – comparison of primary cells and hepatocarcinoma cell line

Abstract

This thesis was conducted with Finnish Hub for Development and Validation of Integrated Approaches (FHAIVE). FHAIVE research group is a part of Faculty of Medicine and Health Technology of Tampere University. The group focuses on replacing animal testing with in vitro models, computing and machine learning.

The EU funded project LiverTarget aims to invent efficient medical treatments to cardiometabolic diseases which are one of the leading causes of death in the world. LiverTarget aims to use RNA inhibition to reduce the expression of inflammatory genes which could work as a treatment to these diseases. The aim of the thesis was to define an experimental setup for the research group’s LiverTarget project. The objective of the thesis was to culture and compare primary hepatocytes and hepatocarcinoma cell line and their metabolic activity. This was done by studying their metabolic activity with commercial kits that measure secretion of urea and albumin and by measuring expression of genes typical to liver and important to metabolism with reverse transcriptase quantitative polymerase chain reaction.

The results show that the primary cells maintain their liver-like functions better than the hepatocarcinoma cell line. Urea production was higher in the primary cells. Also, the gene that encodes the production of albumin, ALB, and urea linked gene ARG1 were more expressed in the primary cells which indicates towards higher secretion. The gene expression was higher in the primary cells in most of all the other tested genes as well.

The results of this thesis will be used in the implementation of the LiverTarget experiments. The test setup was mostly applicable but will need some improvements before the actual project experiments begin. In the future, repetitions of the test are needed and a new cell line with characteristics between the primary cells and hepatocarcinoma cells could be added to the comparison.