Cell models - Liver cells
8 important questions on Cell models - Liver cells
In toxicology what are important types of cells? And why (4)
• Both receive large amounts of blood
• Both harbour large variety of metabolizing enzymes (phase I and phase II)
• Both polarized cells, both have a number of specific transporters
• Involved in elimination of compounds via urine or bile
Give some basic knowledge about the kidney cells.
- Two main classes of cells: hepatocytes and non parenchymal cells
- Sinussoidal endothelial cells: tox and regeneration
- Kupffer cells: tox?
- Hepatic stellate cells: involved in repair
What are the available hepatocyte cell lines?(5)
Limitation?
• Hep3B
• Huh7 CyP3A4 metabolism
• Fa2N4
• HepaRG
Come from cancer cells -> low metabolizing enzymes
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Give an example of why CYP is needed for toxicity testing.
Needs to be metabolized to be toxic.
We can have different types of cultures of primary hepatocytes. What types?
- Freshly isolated: loose CYP activity rapidly.
- Collagen coating
- Sandwich culture: between two layers of collagen. Polarization possible. Basolateral side and a apical side.
- Spheroid: Hanging drop, cells produce ECM closely resembled to liver
What type of culture resemble the liver the most? Why? (4)
ð Multiple primary human hepatocytes (form males and female)
ð Cocultures possible with Kupffer cells and SECs
ð 28 days viability (normally 7 days)
ð Mimics cell density in the liver
IPSC-derived hepatocytes. What do you need to do to generate them?
forkhead box (FOX)A3
and Prospero homeobox protein (PROX1) “HC3X“ induceable by doxycycline (day 4 to 20)
What is important for iPSC hepatocytes?
Improved media with enriched non-essential amino acids (5x higher), 2% glycine
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