Bioengineering Liver Transplantation II

A special issue of Bioengineering (ISSN 2306-5354).

Deadline for manuscript submissions: closed (30 September 2022) | Viewed by 22997

Special Issue Editors


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Guest Editor
Department of Surgery, Erasmus MC Transplant Institute, University Medical Center Rotterdam, 3015 GD Rotterdam, The Netherlands
Interests: liver extracellular matrix proteins; recellularization; biomechanical analysis; organoids; bile duct regeneration
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Guest Editor
Department of Surgery, Erasmus MC-University Medical Center, 3015 CE Rotterdam, The Netherlands
Interests: regenerative medicine; liver transplantation; stem cells; tissue engineering; liver organoids
Special Issues, Collections and Topics in MDPI journals
Department of Clinical Sciences, Faculty of Veterinary Medicine, Regenerative Medicine Center Utrecht, Utrecht University, Uppsalalaan 8, 3584 CT Utrecht, The Netherlands
Interests: bioprinting; liver iPSC; organoids; tissue engineering; regenerative medicine
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

The aim of this second Special Issue “Bioengineering Liver Transplantation—Part II” is to understand, review, and evaluate new and exciting opportunities from the field on regenerative medicine, biomaterials, and stem cell research for bioengineering human liver grafts that can be applied for transplantation and personalized treatment of end-stage liver disease.

The development of culture conditions for long-term expansion of liver organoids and induced pluripotent stem cells (iPSC), including novel protocols for hepatocyte and cholangiocyte differentiation, is moving forward, fostering their use in liver tissue engineering.

In this Special Issue, we again welcome reviews and original papers focusing on hepatic cell sources, including adult hepatic stem cells, organoids, fetal and induced pluripotent stem cells, and primary cells (i.e., hepatocytes, cholangiocytes, and endothelial cells) and how these cells can be applied in tissue engineering strategies to generate implantable and personalized liver grafts. Potential topics include but are not limited to liver tissue engineering, liver regeneration, graft repair, liver stem cells and organoids, bioscaffolds, and 3D printing.

We invite you to contribute original research papers, as well as comprehensive reviews, aligned with these themes, to advance and improve the actual state of the art in liver bioengineering and provide new opportunities to solve the imminent medical problem of organ and tissue shortage for transplantation.

Dr. Monique M. A. Verstegen
Prof. Dr. Luc J. W. van der Laan
Dr. Bart Spee
Guest Editors

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Keywords

  • liver regeneration
  • liver stem cells and organoids
  • liver tissue engineering
  • graft repair
  • bioscaffolds
  • bioprinting

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Published Papers (4 papers)

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Research

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17 pages, 3819 KiB  
Article
Establishment of a Serum-Free Hepatocyte Cryopreservation Process for the Development of an “Off-the-Shelf” Bioartificial Liver System
by Ji-Hyun Lee, Hey-Jung Park, Young-A Kim, Doo-Hoon Lee, Jeong-Kwon Noh, Jong-Gab Jung, Hee-Hoon Yoon, Suk-Koo Lee and Sanghoon Lee
Bioengineering 2022, 9(12), 738; https://doi.org/10.3390/bioengineering9120738 - 29 Nov 2022
Cited by 4 | Viewed by 2747
Abstract
To use hepatocytes immediately when necessary for hepatocyte transplantation and bioartificial liver (BAL) systems, a serum-free cryopreservation protocol ensuring the high survival of hepatocytes and maintenance of their functions should be developed. We established a serum-free protocol for the cryopreservation of primary hepatocytes, [...] Read more.
To use hepatocytes immediately when necessary for hepatocyte transplantation and bioartificial liver (BAL) systems, a serum-free cryopreservation protocol ensuring the high survival of hepatocytes and maintenance of their functions should be developed. We established a serum-free protocol for the cryopreservation of primary hepatocytes, hepatocyte spheroids, and hepatocyte spheroid beads in liquid nitrogen. The serum-free cryopreservation solutions showed a significantly higher performance in maintaining enhanced viability and ammonia removal, urea secretion, and the albumin synthesis of hepatocyte spheroids and spheroid beads. The serum-free thawing medium, containing human serum albumin (HSA) and N-acetylcysteine (NAC), was compared with a fetal bovine serum-containing thawing medium for the development of a serum-free thawing medium. Our results show that hepatocyte spheroids and spheroid beads thawed using a serum-free thawing medium containing HSA and NAC exhibited increased hepatocyte viability, ammonia removal, urea secretion, and albumin synthesis compared to those thawed using the serum-containing medium. Finally, we evaluated the liver functions of the cryopreserved BAL system-applied serum-free cryopreservation process compared to the fresh BAL system. The ammonia removal efficiency of the cryopreserved hepatocyte spheroids BAL was lower than or similar to that of the fresh BAL system. Additionally, the urea concentrations in the media of all three BAL systems were not significantly different during BAL system operation. This cryopreserved spheroid-based BAL system using a serum-free process will be a good candidate for the treatment of patients. Full article
(This article belongs to the Special Issue Bioengineering Liver Transplantation II)
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19 pages, 2834 KiB  
Article
Normothermic Ex Vivo Liver Platform Using Porcine Slaughterhouse Livers for Disease Modeling
by Melanie Krüger, Alicia Ruppelt, Benjamin Kappler, Elke Van Soest, Roos Anne Samsom, Guy C. M. Grinwis, Niels Geijsen, J. Bernd Helms, Marco Stijnen, Linda M. Kock, Marco Rasponi, Hans S. Kooistra and Bart Spee
Bioengineering 2022, 9(9), 471; https://doi.org/10.3390/bioengineering9090471 - 14 Sep 2022
Cited by 2 | Viewed by 3245
Abstract
Metabolic and toxic liver disorders, such as fatty liver disease (steatosis) and drug-induced liver injury, are highly prevalent and potentially life-threatening. To allow for the study of these disorders from the early stages onward, without using experimental animals, we collected porcine livers in [...] Read more.
Metabolic and toxic liver disorders, such as fatty liver disease (steatosis) and drug-induced liver injury, are highly prevalent and potentially life-threatening. To allow for the study of these disorders from the early stages onward, without using experimental animals, we collected porcine livers in a slaughterhouse and perfused these livers normothermically. With our simplified protocol, the perfused slaughterhouse livers remained viable and functional over five hours of perfusion, as shown by hemodynamics, bile production, indocyanine green clearance, ammonia metabolism, gene expression and histology. As a proof-of-concept to study liver disorders, we show that an infusion of free fatty acids and acetaminophen results in early biochemical signs of liver damage, including reduced functionality. In conclusion, the present platform offers an accessible system to perform research in a functional, relevant large animal model while avoiding using experimental animals. With further improvements to the model, prolonged exposure could make this model a versatile tool for studying liver diseases and potential treatments. Full article
(This article belongs to the Special Issue Bioengineering Liver Transplantation II)
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Review

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17 pages, 2058 KiB  
Review
Primary Hepatocyte Isolation and Cultures: Technical Aspects, Challenges and Advancements
by Impreet Kaur, Ashwini Vasudevan, Preety Rawal, Dinesh M. Tripathi, Seeram Ramakrishna, Savneet Kaur and Shiv K. Sarin
Bioengineering 2023, 10(2), 131; https://doi.org/10.3390/bioengineering10020131 - 18 Jan 2023
Cited by 17 | Viewed by 11487
Abstract
Hepatocytes are differentiated cells that account for 80% of the hepatic volume and perform all major functions of the liver. In vivo, after an acute insult, adult hepatocytes retain their ability to proliferate and participate in liver regeneration. However, in vitro, prolonged culture [...] Read more.
Hepatocytes are differentiated cells that account for 80% of the hepatic volume and perform all major functions of the liver. In vivo, after an acute insult, adult hepatocytes retain their ability to proliferate and participate in liver regeneration. However, in vitro, prolonged culture and proliferation of viable and functional primary hepatocytes have remained the major and the most challenging goal of hepatocyte-based cell therapies and liver tissue engineering. The first functional cultures of rat primary hepatocytes between two layers of collagen gel, also termed as the “sandwich cultures”, were reported in 1989. Since this study, several technical developments including choice of hydrogels, type of microenvironment, growth factors and culture conditions, mono or co-cultures of hepatocytes along with other supporting cell types have evolved for both rat and human primary hepatocytes in recent years. All these improvements have led to a substantial improvement in the number, life-span and hepatic functions of these cells in vitro for several downstream applications. In the current review, we highlight the details, limitations and prospects of different technical strategies being used in primary hepatocyte cultures. We discuss the use of newer biomaterials as scaffolds for efficient culture of primary hepatocytes. We also describe the derivation of mature hepatocytes from other cellular sources such as induced pluripotent stem cells, bone marrow stem cells and 3D liver organoids. Finally, we also explain the use of perfusion-based bioreactor systems and bioengineering strategies to support the long-term function of hepatocytes in 3D conditions. Full article
(This article belongs to the Special Issue Bioengineering Liver Transplantation II)
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14 pages, 1213 KiB  
Review
Bioengineering Liver Organoids for Diseases Modelling and Transplantation
by Junzhi Li, Jing Chu, Vincent Chi Hang Lui, Shangsi Chen, Yan Chen and Paul Kwong Hang Tam
Bioengineering 2022, 9(12), 796; https://doi.org/10.3390/bioengineering9120796 - 13 Dec 2022
Cited by 7 | Viewed by 4568
Abstract
Organoids as three-dimension (3D) cellular organizations partially mimic the physiological functions and micro-architecture of native tissues and organs, holding great potential for clinical applications. Advances in the identification of essential factors including physical cues and biochemical signals for controlling organoid development have contributed [...] Read more.
Organoids as three-dimension (3D) cellular organizations partially mimic the physiological functions and micro-architecture of native tissues and organs, holding great potential for clinical applications. Advances in the identification of essential factors including physical cues and biochemical signals for controlling organoid development have contributed to the success of growing liver organoids from liver tissue and stem/progenitor cells. However, to recapitulate the physiological properties and the architecture of a native liver, one has to generate liver organoids that contain all the major liver cell types in correct proportions and relative 3D locations as found in a native liver. Recent advances in stem-cell-, biomaterial- and engineering-based approaches have been incorporated into conventional organoid culture methods to facilitate the development of a more sophisticated liver organoid culture resembling a near to native mini-liver in a dish. However, a comprehensive review on the recent advancement in the bioengineering liver organoid is still lacking. Here, we review the current liver organoid systems, focusing on the construction of the liver organoid system with various cell sources, the roles of growth factors for engineering liver organoids, as well as the recent advances in the bioengineering liver organoid disease models and their biomedical applications. Full article
(This article belongs to the Special Issue Bioengineering Liver Transplantation II)
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