Preservation of Cell Structure, Metabolism, and Biotransformation Activity of Liver‐On‐Chip Organ Models by Hypothermic Storage

Abstract

The liver is a central organ in the metabolization of nutrition, endogenous and exogenous substances, and xenobiotic drugs. The emerging organ-on-chip technology has paved the way to model essential liver functions as well as certain aspects of liver disease in vitro in liver-on-chip models. However, a broader use of this technology in biomedical research is limited by a lack of protocols that enable the short-term preservation of preassembled liver-on-chip models for stocking or delivery to researchers outside the bioengineering community. For the first time, this study tested the ability of hypothermic storage of liver-on-chip models to preserve cell viability, tissue morphology, metabolism and biotransformation activity. In a systematic study with different preservation solutions, liver-on-chip function can be preserved for up to 2 d using a derivative of the tissue preservation solution TiProtec, containing high chloride ion concentrations and the iron chelators LK614 and deferoxamine, supplemented with polyethylene glycol (PEG). Hypothermic storage in this solution represents a promising method to preserve liver-on-chip function for at least 2 d and allows an easier access to liver-on-chip technology and its versatile and flexible use in biomedical research.

An Overview of Biotransformation and Toxicity of Diterpenes

“…Some biocatalysts models to study biotransformation in vitro include microorganisms, such as bacteria, filamentous fungi, yeasts, plant cells, and microalgae [ 34 ]. Furthermore, intestinal and liver models such as immortalized cell lines (e.g., Caco-2 and HT-29) [ 35 ], intestinal organoids [ 35 ], isolated perfused liver [ 33 ], enzyme preparations (e.g., liver microsomes, cytosolic and S9 fractions) [ 33 ], membrane biohybrid systems of human hepatocytes [ 36 ], primary human hepatocytes with membrane bioreactors [ 36 ], and liver-on-chip models [ 37 ] have been also used.…”

Nevirapine Biotransformation Insights: An Integrated In Vitro Approach Unveils the Biocompetence and Glutathiolomic Profile of a Human Hepatocyte-Like Cell 3D Model

“…In particular, the organ-on-a-chip technology emerged as promising in vitro models by offering the possibility of mimicking blood circulation by creating a laminar fluid flow, seeding high cell densities, mimicking tissue architecture in a three-dimensional fashion, consuming low cell and reactant amounts and allowing interconnected culture of different cell types [24,25]. However, until today, few authors performed a thorough study on drug metabolism in such devices [26,27]. One of the biggest challenges of using organ-on-a-chip technology is still the high adsorption of small molecules to the microfluidic device materials (e.g., PDMS) requiring additional studies to allow the comparison with already existing data generated using polystyrene as material [26,28].…”

Advanced Fabrication Techniques of Microengineered Physiological Systems

“…Injection molding is predominantly used during large-scale fabrication productions and focused on developing only the first element of OOCs (e.g., the microfluidic device) [58]. Injection molding was used to fabricate a liver-on-a-chip to study hypothermic storage [52]. While injection molding appears to be a simple and easy process to use, it requires familiarity and experience with it to be successfully performed at the micro level [58].…”