Tissue-Specific Microparticles Improve Organoid Microenvironment for Efficient Maturation of Pluripotent Stem-Cell-Derived Hepatocytes

Abstract

Liver organoids (LOs) are receiving considerable attention for their potential use in drug screening, disease modeling, and transplantable constructs. Hepatocytes, as the key component of LOs, are isolated from the liver or differentiated from pluripotent stem cells (PSCs). PSC-derived hepatocytes are preferable because of their availability and scalability. However, efficient maturation of the PSC-derived hepatocytes towards functional units in LOs remains a challenging subject. The incorporation of cell-sized microparticles (MPs) derived from liver extracellular matrix (ECM), could provide an enriched tissue-specific microenvironment for further maturation of hepatocytes inside the LOs. In the present study, the MPs were fabricated by chemical cross-linking of a water-in-oil dispersion of digested decellularized sheep liver. These MPs were mixed with human PSC-derived hepatic endoderm, human umbilical vein endothelial cells, and mesenchymal stromal cells to produce homogenous bioengineered LOs (BLOs). This approach led to the improvement of hepatocyte-like cells in terms of gene expression and function, CYP activities, albumin secretion, and metabolism of xenobiotics. The intraperitoneal transplantation of BLOs in an acute liver injury mouse model led to an enhancement in survival rate. Furthermore, efficient hepatic maturation was demonstrated after ex ovo transplantation. In conclusion, the incorporation of cell-sized tissue-specific MPs in BLOs improved the maturation of human PSC-derived hepatocyte-like cells compared to LOs. This approach provides a versatile strategy to produce functional organoids from different tissues and offers a novel tool for biomedical applications.

Mimicking Native Liver Lobule Microarchitecture In Vitro with Parenchymal and Non-parenchymal Cells Using 3D Bioprinting for Drug Toxicity and Drug Screening Applications

“…The hepatotoxicity prediction in existing liver systems was limited due to the lack of native microenvironmental cues, hepatocyte polarity, and metabolic functions. ,, In vitro liver models encompassing liver ECM preserved significant ECM components, including growth factors, cytokines, and chemokines that in turn aided in maintaining an aggregate culture, polarity, zonation, and metabolic competence of hepatocytes. ,, An ideal bioink hydrogel providing heterotypic cellular interactions, cell–matrix interactions, and mimics tissue-specific physiological conditions is critical during bioprinting of cell-laden liver constructs. ,, Herein, we fabricated liver ECM-based bioink for cell encapsulation, with bulking agent gelatin and bioactive silk fibroin to augment the rheological behavior, post-printing features, and liver cell functionality . Thus, the inclusion of silk fibroin in bioink not only acted as a reinforcing supportive material, indeed provided active biological cues for enhanced cellular attachment through integrin mediated pathway. , Aiming to develop liver ECM-based bioink, a co-printable basic liver ECM bioink composition was optimized as 7% (w/v) gelatin, 1% (w/v) BM, 1% (w/v) AA, and 1% (w/v) liver ECM (Figure ).…”

“…9,16,41 In vitro liver models encompassing liver ECM preserved significant ECM components, including growth factors, cytokines, and chemokines that in turn aided in maintaining an aggregate culture, polarity, zonation, and metabolic competence of hepatocytes. 22,42,43 An ideal bioink hydrogel providing heterotypic cellular interactions, cell−matrix interactions, and mimics tissue-specific physiological conditions is critical during bioprinting of cell-laden liver constructs. 22,23,44 Herein, we fabricated liver ECM-based bioink for cell encapsulation, with bulking agent gelatin and bioactive silk fibroin to augment the rheological behavior, post-printing features, and liver cell functionality.…”

Challenges for the Applications of Human Pluripotent Stem Cell-Derived Liver Organoids

“…ECM plays an important role in cell differentiation and organogenesis and may improve hepatic maturation of PSC-LOs. Zahmatkesh et al (2021) efficiently improved hepatic maturation of LOs by mixing microparticles made of decellularized liver matrix with human PSC-derived endoderm cells, mesenchymal stromal cells, and HUVECs.…”

Modeling Hepatotropic Viral Infections: Cells vs. Animals

“…The HBV infection decreased the expression of hepatic-specific genes and increased the concentration of early biomarkers for acute liver failure, alanine aminotransferase (ALT), and lactate dehydrogenase (LDH), in the supernatant of infected organoids. The iPSCs -derived 3D liver organoids had the advantage of providing the HBV infection models for precision medicine [ 85 ]. Crignis and colleagues developed liver organoids from healthy donors and HBV-infected patients.…”