Primary Hepatocytes Cultured on a Fiber-Embedded PDMS Chip to Study Drug Metabolism.

Yaowen Liu,Ke Hu,Yihao Wang

doi:10.3390/polym9060215

Yaowen Liu, Ke Hu + Show 1 more

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https://doi.org/10.3390/polym9060215

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Abstract

In vitro drug screening using reliable and predictable liver models remains a challenge. The identification of an ideal biological substrate is essential to maintain hepatocyte functions during in vitro culture. Here, we developed a fiber-embedded polydimethylsiloxane (PDMS) chip to culture hepatocytes. Hepatocyte spheroids formed in this device were subjected to different flow rates, of which a flow rate of 50 μL/min provided the optimal microenvironment for spheroid formation, maintained significantly higher rates of albumin and urea synthesis, yielded higher CYP3A1 (cytochrome P450 3A1) and CYP2C11 (cytochrome P450 2C11) enzyme activities for metabolism, and demonstrated higher expression levels of liver-specific genes. In vitro metabolism tests on tolbutamide and testosterone by hepatocytes indicated predicted clearance rates of 1.98 ± 0.43 and 40.80 ± 10.13 mL/min/kg, respectively, which showed a good in vitro–in vivo correspondence. These results indicate that this system provides a strategy for the construction of functional engineered liver tissue that can be used to study drug metabolism.

Highlights

The liver is the major organ for drug biotransformation
These results indicate that this system provides a strategy for the construction of functional engineered liver tissue that can be used to study drug metabolism
The results showed that the flow- and fiber-based microfluidic chip system could achieve longer hepatocyte culture, a higher level of hepatocyte functions, and more accurate prediction of drug metabolism compared to the available methods

Summary

Introduction

In vitro drug screening is the primary alternative to animal testing due to fast, cheap, and safe drug development, with minimalized testing on animals. Numerous in vitro models have been proposed, such as liver slices, primary cells, and liver cell lines; they all have various disadvantages. Liver slices exhibit a limited lifespan in culture (approximately 1 week), limiting their widespread use in vitro [1]. Liver cell lines, such as HepG2, have no or very low levels of important drug-metabolizing enzymes and transporters [2]. To improve the predictive abilities of primary hepatocytes, novel strategies need to be developed to maintain their morphology and function in vitro

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