Advanced Techniques and Awaited Clinical Applications for Human Pluripotent Stem Cell Differentiation into Hepatocytes

Abstract

Liver transplantation is currently the only curative treatment for several liver diseases such as acute liver failure, end-stage liver disorders, primary liver cancers and certain genetic conditions.Unfortunately, despite improvements to transplantation techniques, including live donor transplantation, the number of organs available remains insufficient to meet patient needs.Hepatocyte transplantation has enabled some encouraging results as an alternative to organ transplantation, but primary hepatocytes are little available and cannot be amplified using traditional 2D culture systems. Indeed, although recent studies have tended to show that 3D culture enables long term hepatocyte culture, it is still agreed that like most adult primary cell types, they remain refractory to in vitro expansion. Because of their exceptional properties, human pluripotent stem cells (hPSCs) can be amplified indefinitely and differentiated into any cell type, including liver cells. While many teams have worked on hepatocyte differentiation, there has been a consensus that cells obtained after hPSC differentiation have more foetal than adult hepatocyte characteristics. New technologies have been used to improve the differentiation process in recent years. This review discusses the technical improvements made to hepatocyte differentiation protocols and the clinical approaches developed to date and anticipated in the near future.

Induction and Maturation of Hepatocyte-Like Cells In Vitro: Focus on Technological Advances and Challenges

“…However, the costs and complex technology involved in establishing and maintaining organoid cultures are causes of the limited research. In addition, the batch effect caused by varying environments and culture durations affect the experimental results (Luce et al, 2021) (Figure 4).…”

Evidence of Adult Features and Functions of Hepatocytes Differentiated from Human Induced Pluripotent Stem Cells and Self-Organized as Organoids

“…Through guided differentiation, human-induced pluripotent stem cells (hiPSCs) offer a reliable source of hepatocytes that are certainly proving themselves useful in numerous tissue engineering and personalized medicine applications [1][2][3][4][5][6]. However, concerns persist regarding the lack of functional maturity and long-term maintenance in culture of hiPSCderived hepatocytes (iHeps).…”

“…Indeed, iHeps still display a fetal hepatic phenotype rather than an adult one [7,8] and, in particular, they continue to express fetal markers such as α-fetoprotein (AFP), as well as being poor at reproducing key mature functions, such as the activity of many detoxification enzymes [8,9]. Improved protocols and 3D culture systems have shown that the hepatic functions of iHeps can be enhanced, thus contributing to the study of liver development, diseases and the efficacy of medicinal products [3][4][5]10,11], but hepatocytes with adult features have still not been generated. This hurdle currently remains the principal drawback of iHeps in terms of applications where a considerable degree of maturity is required, such as pharmacology/toxicology studies and the development of in vitro systems to model liver physiology and pathophysiology.…”

Bioprinting of a Hepatic Tissue Model Using HumanInduced Pluripotent Stem Cell-derived Hepatocytes for Drug-Induced Hepatotoxicity Evaluation

“…Nonetheless, hiPSCs could be a promising renewable and easily accessible cell source for the generation of functional hepatocytes which may obtain full maturity with the application of future biotechnologies. When applied in the bioprinting of hepatic tissue models using proper printing techniques and bioinks, hiPSC-derived hepatocytes (hiPSC-Heps) have demonstrated well-maintained cellular phenotypes and biofunctions[ 25 ]. Bioprinting of hiPSCs and human embryonic stem cells was first reported in 2015 by Faulkner-Jones et al .…”