Abstract
3D (three-dimensional) cultures are considered to be an effective method for toxicological studies; however, little evidence has been reported whether 3D cultures have an impact on hepatocellular physiology regarding lipid or glucose metabolism. In the present study, we conducted physiological characterization of hepatoma cell lines HepG2 and HepaRG cells cultured in 3D conditions using a hanging drop method to verify the effect of culture environment on cellular responses. Apo (Apolipoprotein)B as well as albumin secretion was augmented by 3D cultures. Expression of genes related to not only drug, but also glucose and lipid metabolism were significantly enhanced in 3D cultured HepaRG spheroids. Furthermore, mRNA levels of CYP (cytochrome P450) enzymes following exposure to corresponding inducers increased under the 3D condition. These data suggest that this simple 3D culture system without any special biomaterials can improve liver-specific characteristics including lipid metabolism. Considering that the system enables high-throughput assay, it may become a powerful tool for compound screening concerning hepatocellular responses in order to identify potential drugs.
Highlights
The demand to establish physiological assays has increased, in particular for compound screening and drug development [1,2]
We assessed the effect of 3D cultures on hepatocellular characteristics and validated the hypothesis that 3D cultures can improve levels of apoB secretion per viable cell and mRNA levels of various genes exclusively expressed in the liver
It is noteworthy that this 3D culture system requires neither special biomaterials such as basement membrane proteins nor particular equipment such as bioreactors, thereby the effect of cultural environment change itself can be directly evaluated without any external factors
Summary
The demand to establish physiological assays has increased, in particular for compound screening and drug development [1,2]. Cell-based assays are essential for validating the pharmacological activity of drug candidates; development for a large number of these candidates is often discontinued during subsequent animal testing or clinical trials, which entails significant commitments in terms of costs, time and energy. The cells are assembled by gravity force and migrate with polarity and an extensive extracellular matrix including collagen is formed within the spheroids as in vivo tissues. When it comes to HepG2 spheroids, there are canaliculi-like structures observed by SEM (scanning electron microscope) [14]. Studies on hepatic function, such as lipid metabolism, have not been reported to date
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