A perfusion incubator liver chip for 3D cell culture with application on chronic hepatotoxicity testing

• Published in: Scientific reports Vol. 7; no. 1; pp. 14528 - 16
• Main Authors: Yu, Fang, Deng, Rensheng, Hao Tong, Wen, Huan, Li, Chan Way, Ng, IslamBadhan, Anik, Iliescu, Ciprian, Yu, Hanry
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 06.11.2017; Nature Publishing Group
• Subjects: 13/106; 13/107; 13/62; 631/154/1435; 639/166/985; Acetaminophen; Animals; Biosafety; Carbon dioxide; Cell culture; Cell Culture Techniques - methods; Cell Survival; Contamination; Culture media; Diclofenac; Drug dosages; Enzymatic activity; Gas Chromatography-Mass Spectrometry; Hepatocytes; Hepatocytes - cytology; Hepatocytes - drug effects; Hepatotoxicity; Humanities and Social Sciences; Lab-On-A-Chip Devices; Liver; Liver - cytology; Liver - drug effects; Male; Mass transfer; Mechanical stimuli; multidisciplinary; Perfusion; Perfusion - instrumentation; Perfusion - methods; pH effects; Rats; Rats, Wistar; Risk reduction; Science; Science (multidisciplinary); Shear stress; Spheroids; Sterility; Toxicity Tests, Chronic - instrumentation; Toxicity Tests, Chronic - methods; Urea
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-017-13848-5

Liver chips have been developed to recapitulate in vivo physiological conditions to enhance hepatocyte functions for assessing acute responses to drugs. To develop liver chips that can assess repeated dosing chronic hepatotoxicity, we need to ensure that hepatocyte functions be maintained at constant values over two weeks in stable culture conditions of sterility, temperature, pH, fluidic-flow of culture media and drugs. We have designed a perfusion-incubator-liver-chip (PIC) for 3D cell culture, that assures a tangential flow of the media over the spheroids culture. Rat hepatocyte spheroids constrained between a cover glass and a porous-ultrathin Parylene C membrane experienced optimal mass transfer and limited shear stress from the flowing culture media; maintained cell viability over 24 days. Hepatocyte functions were significantly improved and maintained at constant values (urea, albumin synthesis, and CYP450 enzyme activities) for 14 days. The chip act as an incubator, having 5% CO 2 pressure-driven culture-media flow, on-chip heater and active debubbler. It operates in a biosafety cabinet, thus minimizing risk of contamination. The chronic drug response to repeated dosing of Diclofenac and Acetaminophen evaluated in PIC were more sensitive than the static culture control.