Introduction
Animal models and two-dimensional (2D) cell cultures frequently fail to replicate human drug metabolism and liver toxicity in vivo, therefore there is a need for 3D Culture of HepaRG cells. Indeed, there is only a 30 to 50% concordance between animal and human models when it comes to hepatic drug toxicity. Furthermore, primary hepatocytes cultivated in 2D lose their functions quickly and cannot be used to examine pharmacological effects over time.
Their microfluidic chip was created to resemble the microstructure of the liver and to allow for more physiological culture conditions than earlier devices. Indeed, in addition to attaining 3D hepatocyte culture, the cells’ architecture allowed them to spatially organize into hepatocyte cords with in vivo-like dimensions.
How to culture vascularized & immunocompetent 3D models in a standard Multiwell
Abstract
The authors state that “The development of livers-on-a-chip aims to provide pharmaceutical companies with reliable systems to perform drug screening and toxicological studies. To that end, microfluidic systems are engineered to mimic the functions and architecture of this organ.
In this context we have designed a device that reproduces a series of liver microarchitectures, each permitting the 3D culture of hepatocytes by confining them to a chamber that is separated from the medium conveying channel by very thin slits.
We modified the structure to ensure its compatibility with the culture of hepatocytes from different sources. Our device was adapted to the migratory and adhesion properties of the human HepaRG cell line at various stages of differentiation. Using this device, it was possible to keep the cells alive for more than 14 days, during which they achieved a 3D organization and acquired or maintained their differentiation into hepatocytes.
Albumin secretion, as well as functional bile canaliculi, were confirmed on the liver-on-a-chip. Finally, an acetaminophen toxicological assay was performed. With its multiple micro-chambers for hepatocyte culture, this microfluidic device architecture offers a promising opportunity to provide new tools for drug screening applications.”
References
Boul M, Benzoubir N, Messina A, Ghasemi R, Mosbah IB, Duclos-Vallée JC, Dubart-Kupperschmitt A, Le Pioufle B. A versatile microfluidic tool for the 3D culture of HepaRG cells seeded at various stages of differentiation. Sci Rep. 2021 Jul 7;11(1):14075. doi: 10.1038/s41598-021-92011-7. PMID: 34234159; PMCID: PMC8263583.
FAQ
A need for 3D culture models like those for HepaRG cells exists. Animal models and 2D cell cultures often do not successfully replicate human drug metabolism. They also fail to show liver toxicity as it occurs in humans. A low concordance rate of only 30 to 50 percent is seen between animal and human models regarding hepatic drug toxicity. Furthermore, when primary hepatocytes are cultivated in 2D, their functions are lost quickly. This loss of function means they cannot be used to examine the effects of pharmaceuticals over time. New models are required to address these limitations.
The development of "livers-on-a-chip" is intended to provide reliable systems to pharmaceutical companies. These systems are meant to be used for performing drug screening. They are also used for toxicological studies. Microfluidic systems are engineered for this purpose. They are designed to copy the functions and architecture of the actual organ. This new device is one such system, designed to reproduce a series of liver microarchitectures. The overall goal is to create more accurate tools for testing. These tools could improve the drug development process by offering better predictions of human responses.
This microfluidic chip was designed to resemble the microstructure of the liver. It allows for more physiological culture conditions than were possible in earlier devices. A series of liver microarchitectures are reproduced by the device. These permit the 3D culture of hepatocytes. The cells are confined to a chamber. This chamber is separated by very thin slits from the channel that conveys the medium. The device’s structure was modified. This was done to ensure it is compatible with hepatocytes from different sources. For instance, the device was adapted for the migratory and adhesion properties of the human HepaRG cell line at various stages of differentiation.
Using this device, cells were kept alive for more than 14 days. A 3D organisation was achieved by the cells during this period. They also acquired or maintained their differentiation into hepatocytes. The architecture of the cells allowed for spatial organisation. They formed hepatocyte cords with dimensions similar to those found in vivo. Key liver functions were also confirmed. Albumin secretion was observed in the liver-on-a-chip. Functional bile canaliculi were also confirmed. To test its application, an acetaminophen toxicological assay was performed. The device, with its multiple micro-chambers, is seen as a promising tool for drug screening.
