3D Cultivation Techniques for Primary Human Hepatocytes

Abstract

One of the main challenges in drug development is the prediction of in vivo toxicity based on in vitro data. The standard cultivation system for primary human hepatocytes is based on monolayer cultures, even if it is known that these conditions result in a loss of hepatocyte morphology and of liver-specific functions, such as drug-metabolizing enzymes and transporters. As it has been demonstrated that hepatocytes embedded between two sheets of collagen maintain their function, various hydrogels and scaffolds for the 3D cultivation of hepatocytes have been developed. To further improve or maintain hepatic functions, 3D cultivation has been combined with perfusion. In this manuscript, we discuss the benefits and drawbacks of different 3D microfluidic devices. For most systems that are currently available, the main issues are the requirement of large cell numbers, the low throughput, and expensive equipment, which render these devices unattractive for research and the drug-developing industry. A higher acceptance of these devices could be achieved by their simplification and their compatibility with high-throughput, as both aspects are of major importance for a user-friendly device.

Three‐dimensional cell culture models for anticancer drug screening: Worth the effort?

“…Hydrogels derived from a natural origin are highly bioactive due to the presence of a wide variety of endogenous molecules, which can be advantageous for many cellular processes including proliferation or differentiation. One of the most commonly used naturally‐derived hydrogels is the so‐called Matrigel, which is composed of a mixture of ECM proteins secreted by Engelbreth‐Holm‐Swarm sarcoma cells (Bachmann et al, ). However, such scaffolds are often not well characterized making it challenging to identify the endogenous factors that promote a certain cell behavior.…”

Application of Micropatterned Cocultured Hepatocytes to Evaluate the Inductive Potential and Degradation Rate of Major Xenobiotic Metabolizing Enzymes

“…unattractive for the pharmaceutical industry (Bachmann et al, 2015). The HepatoPac coculture model consists of primary hepatocytes that are micropatterned to form a discrete microarchitecture or "hepatocyte islands" that are surrounded by supporting fibroblasts resulting in longterm viability and metabolic function for several weeks (Khetani and Bhatia, 2008).…”

Decellularized caprine liver extracellular matrix as a 2D substrate coating and 3D hydrogel platform for vascularized liver tissue engineering

“…Two‐dimensional culturing of the hepatocytes is not preferred as they lose their differentiated state over time. Many reports have already showcased the importance of 3D cell culture for long‐term culture of hepatocytes (Bachmann et al, ; Edmondson, Broglie, Adcock, & Yang, ). Thus, in this study, we have also evaluated CLECM‐based hydrogels for the 3D culture of HepG2 cells.…”

“…Direct transplantation of the hepatocytes has been reported with various complications including transient hemodynamic instability during cell infusion, lower survival rates of hepatocytes post‐infusion, lack of proper cellular attachment and host tissue integration, and requirement of repeated cell infusion (Palakkan et al, ). To address these limitations, tissue engineering approaches have come up as a superior alternative (Bachmann et al, ).…”