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

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.

[1]  R. Moghadasali,et al.  Promoting maturation of human pluripotent stem cell-derived renal micro-tissue by incorporation of endothelial and mesenchymal cells. , 2021, Stem cells and development.

[2]  T. Mahmoudi,et al.  Evolution of organoid technology: Lessons learnt in Co-Culture systems from developmental biology. , 2021, Developmental biology.

[3]  H. Baharvand,et al.  Generation of Scalable Hepatic Micro-Tissues as a Platform for Toxicological Studies , 2020, Tissue Engineering and Regenerative Medicine.

[4]  R. Riley,et al.  The evolution of strategies to minimise the risk of human drug-induced liver injury (DILI) in drug discovery and development , 2020, Archives of Toxicology.

[5]  A. Rizvanov,et al.  Cell Culture Based in vitro Test Systems for Anticancer Drug Screening , 2020, Frontiers in Bioengineering and Biotechnology.

[6]  Tiago G Fernandes,et al.  Production of Human Pluripotent Stem Cell-Derived Hepatic Cell Lineages and Liver Organoids: Current Status and Potential Applications , 2020, Bioengineering.

[7]  S. Akbari,et al.  Next-Generation Liver Medicine Using Organoid Models , 2019, Front. Cell Dev. Biol..

[8]  H. Baharvand,et al.  Generation of Transplantable Three-Dimensional Hepatic-Patch to Improve the Functionality of Hepatic Cells in vitro and in vivo. , 2019, Stem cells and development.

[9]  B. Davidson,et al.  Development and characterisation of a Porcine Liver Scaffold. , 2019, Stem cells and development.

[10]  Nicola Elvassore,et al.  Extracellular matrix hydrogel derived from decellularized tissues enables endodermal organoid culture , 2019, Nature Communications.

[11]  H. Järveläinen,et al.  Extracellular matrix-cell interactions: Focus on therapeutic applications. , 2019, Cellular signalling.

[12]  E. Raschi,et al.  Strategies for Early Prediction and Timely Recognition of Drug-Induced Liver Injury: The Case of Cyclin-Dependent Kinase 4/6 Inhibitors , 2019, Front. Pharmacol..

[13]  B. Lin,et al.  Engineered Liver-On-A-Chip Platform to Mimic Liver Functions and Its Biomedical Applications: A Review , 2019, Micromachines.

[14]  Mengjun Dai,et al.  Generation of hepatic spheroids using human hepatocyte-derived liver progenitor-like cells for hepatotoxicity screening , 2019, Theranostics.

[15]  T. He,et al.  A simplified 3D liver microsphere tissue culture model for hepatic cell signaling and drug-induced hepatotoxicity studies , 2019, International journal of molecular medicine.

[16]  R. Cummings,et al.  Generation of fully functional hepatocyte-like organoids from human induced pluripotent stem cells mixed with Endothelial Cells , 2019, Scientific Reports.

[17]  Pere Roca-Cusachs,et al.  Integrins as biomechanical sensors of the microenvironment , 2019, Nature Reviews Molecular Cell Biology.

[18]  C. Nonaka,et al.  Advances in Hepatic Tissue Bioengineering with Decellularized Liver Bioscaffold , 2019, Stem cells international.

[19]  B. Sarmento,et al.  Development and Characterization of Chitosan Microparticles-in-Films for Buccal Delivery of Bioactive Peptides , 2019, Pharmaceuticals.

[20]  G. Daley,et al.  Induced pluripotent stem cells in disease modelling and drug discovery , 2019, Nature reviews genetics.

[21]  H. Baharvand,et al.  Human cardiomyocytes undergo enhanced maturation in embryonic stem cell-derived organoid transplants. , 2019, Biomaterials.

[22]  P. Nell.  Highlight report: spheroids from stem cell-derived hepatocyte-like cells , 2018, Archives of Toxicology.

[23]  Mohammad Kazemi Ashtiani,et al.  Generation of functional human pancreatic organoids by transplants of embryonic stem cell derivatives in a 3D‐printed tissue trapper , 2018, Journal of cellular physiology.

[24]  Weimin Li,et al.  Native-mimicking in vitro microenvironment: an elusive and seductive future for tumor modeling and tissue engineering , 2018, Journal of biological engineering.

[25]  R. Shah,et al.  Kidney decellularized extracellular matrix hydrogels: Rheological characterization and human glomerular endothelial cell response to encapsulation. , 2018, Journal of biomedical materials research. Part A.

[26]  Tiago G Fernandes,et al.  Towards Multi-Organoid Systems for Drug Screening Applications , 2018, Bioengineering.

[27]  H. Baharvand,et al.  Three‐dimensional liver‐derived extracellular matrix hydrogel promotes liver organoids function , 2018, Journal of cellular biochemistry.

[28]  Gordana Vunjak-Novakovic,et al.  Organs-on-a-Chip: A Fast Track for Engineered Human Tissues in Drug Development. , 2018, Cell stem cell.

[29]  H. Baharvand,et al.  Developing a Cost-Effective and Scalable Production of Human Hepatic Competent Endoderm from Size-Controlled Pluripotent Stem Cell Aggregates. , 2018, Stem Cells and Development.

[30]  Sigrid A. Langhans.  Three-Dimensional in Vitro Cell Culture Models in Drug Discovery and Drug Repositioning , 2018, Front. Pharmacol..

[31]  A. Lyne,et al.  Mouse decellularised liver scaffold improves human embryonic and induced pluripotent stem cells differentiation into hepatocyte-like cells , 2017, PloS one.

[32]  Yan Jin,et al.  Liver extracellular matrix promotes BM‐MSCs hepatic differentiation and reversal of liver fibrosis through activation of integrin pathway , 2017, Journal of tissue engineering and regenerative medicine.

[33]  Massoud Vosough,et al.  A Quick update from the Past to Current Status of Human Pluripotent Stem Cell-derived Hepatocyte culture systems , 2017 .

[34]  D. Cho,et al.  Precise stacking of decellularized extracellular matrix based 3D cell-laden constructs by a 3D cell printing system equipped with heating modules , 2017, Scientific Reports.

[35]  C. Billington,et al.  Orexin activation counteracts decreases in nonexercise activity thermogenesis (NEAT) caused by high-fat diet , 2017, Physiology & Behavior.

[36]  D. Brenner,et al.  Protective effect of human serum amyloid P on CCl4-induced acute liver injury in mice , 2017, International journal of molecular medicine.

[37]  Haiyan Zhang,et al.  Endothelial cell‐derived matrix promotes the metabolic functional maturation of hepatocyte via integrin‐Src signalling , 2017, Journal of cellular and molecular medicine.

[38]  A. Brivanlou,et al.  Embryoids, organoids and gastruloids: new approaches to understanding embryogenesis , 2017, Development.

[39]  X. Wen,et al.  Scalable Differentiation of Human iPSCs in a Multicellular Spheroid-based 3D Culture into Hepatocyte-like Cells through Direct Wnt/β-catenin Pathway Inhibition , 2016, Scientific Reports.

[40]  K. Christman,et al.  Controlling stem cell behavior with decellularized extracellular matrix scaffolds. , 2016, Current opinion in solid state & materials science.

[41]  Shay Soker,et al.  Functional Maturation of Induced Pluripotent Stem Cell Hepatocytes in Extracellular Matrix—A Comparative Analysis of Bioartificial Liver Microenvironments , 2016, Stem cells translational medicine.

[42]  S. Badylak,et al.  Solubilized liver extracellular matrix maintains primary rat hepatocyte phenotype in-vitro. , 2016, Journal of Biomedical Materials Research. Part A.

[43]  J. Omens,et al.  Evidence for Mechanisms Underlying the Functional Benefits of a Myocardial Matrix Hydrogel for Post-MI Treatment. , 2016, Journal of the American College of Cardiology.

[44]  S. Strom,et al.  The Use of Induced Pluripotent Stem Cells for the Study and Treatment of Liver Diseases , 2016, Current protocols in toxicology.

[45]  Minoru Seki,et al.  Cell-sized condensed collagen microparticles for preparing microengineered composite spheroids of primary hepatocytes. , 2015, Lab on a chip.

[46]  K. Christman,et al.  Fabrication and characterization of injectable hydrogels derived from decellularized skeletal and cardiac muscle. , 2015, Methods.

[47]  Takanori Takebe,et al.  Vascularized and Complex Organ Buds from Diverse Tissues via Mesenchymal Cell-Driven Condensation. , 2015, Cell stem cell.

[48]  O. Lee,et al.  Cryo-chemical decellularization of the whole liver for mesenchymal stem cells-based functional hepatic tissue engineering. , 2014, Biomaterials.

[49]  Takanori Takebe,et al.  Generation of a vascularized and functional human liver from an iPSC-derived organ bud transplant , 2014, Nature Protocols.

[50]  Takanori Takebe,et al.  Vascularized and functional human liver from an iPSC-derived organ bud transplant , 2013, Nature.

[51]  S. Seif-Naraghi,et al.  Injectable extracellular matrix derived hydrogel provides a platform for enhanced retention and delivery of a heparin-binding growth factor. , 2012, Acta Biomaterialia.

[52]  Silviya P Zustiak,et al.  Hydrolytically degradable poly(ethylene glycol) hydrogel scaffolds with tunable degradation and mechanical properties. , 2010, Biomacromolecules.

[53]  R. Fässler,et al.  Genetic and cell biological analysis of integrin outside-in signaling. , 2009, Genes & development.

[54]  H. Baharvand,et al.  Efficient and cost-effective generation of hepatocyte-like cells through microparticle-mediated delivery of growth factors in a 3D culture of human pluripotent stem cells. , 2018, Biomaterials.

[55]  Ibrahim T. Ozbolat,et al.  Challenges in Bio-fabrication of Organoid Cultures. , 2018, Advances in experimental medicine and biology.

[56]  Pearl Moharil,et al.  Biomaterials and cells for cardiac tissue engineering: Current choices. , 2017, Materials Science and Engineering C: Materials for Biological Applications.

[57]  P. Rees,et al.  Reduced Cationic Nanoparticle Cytotoxicity Based on Serum Masking of Surface Potential. , 2016, Journal of biomedical nanotechnology.

[58]  Ashok Kumar,et al.  Biomaterials for liver tissue engineering , 2013, Hepatology International.