Improved rat liver decellularization by arterial perfusion under oscillating pressure conditions

Struecker, Benjamin; Butter, Antje; Hillebrandt, Karl H.; Polenz, Dietrich; Reutzel‐Selke, Anja; Tang, Peter; Lippert, Steffen; Leder, A.; Rohn, Susanne; Geisel, Dominik; Denecke, Timm; Aliyev, Khalid; Jöhrens, Korinna; Raschzok, Nathanael; Neuhaus, P.; Pratschke, Johann; Sauer, Igor M.

doi:10.1002/term.1948

Cited by 57 publications

(67 citation statements)

References 37 publications

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“…However, the exclusive application of this detergent resulted in incomplete decellularization with nuclear material remaining in the scaffold. In most previous studies, a combination of these two detergents was applied [6,8,10,13,14,16,22,24,26,29,35,36]. As a result, no remaining cellular and nuclear material was detected, and the structure of the extracellular components was maintained (fig.…”

Section: The Decellularization Processmentioning

confidence: 99%

“…As the main route for the perfusion, the portal vein was used. Alternatively, the hepatic artery or the vena cava served as perfusion route [14,16,22,26]. To secure the outflow, the hepatic vessels like the hepatic veins and the vena cava inferior were severed before the perfusion started.…”

Section: The Decellularization Processmentioning

confidence: 99%

“…In fundamental studies, rodents such as mice, rats, ferrets and rabbits are used [5,6,7,8,9,10,11,12,13,14,15,16,17,18,19,20,21,22,23,24]. In translational studies, large animals such as pigs and sheep are preferred [25,26,27,28,29,30,31,32].…”

Section: Source Of Organs For Liver Engineeringmentioning

confidence: 99%

“…The time also ranged from less than 3 h [22] up to several days for total perfusion [27,31]. As the main route for the perfusion, the portal vein was used.…”

Section: The Decellularization Processmentioning

confidence: 99%

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Bioengineered Livers: A New Tool for Drug Testing and a Promising Solution to Meet the Growing Demand for Donor Organs

et al. 2016

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Background: Organ engineering is a new innovative strategy to cope with two problems: the need for physiological models for pharmacological research and donor organs for transplantation. A functional scaffold is generated from explanted organs by removing all cells (decellularization) by perfusing the organ with ionic or nonionic detergents via the vascular system. Subsequently the acellular scaffold is reseeded with organ-specific cells (repopulation) to generate a functional organ. Summary: This review gives an overview of the state of the art describing the decellularization process, the subsequent quality assessment, the repopulation techniques and the functional assessment. It emphasizes the use of scaffolds as matrix for culturing human liver cells for drug testing. Further, it highlights the techniques for transplanting these engineered scaffolds in allogeneic or xenogeneic animals in order to test their biocompatibility and use as organ grafts. Key Messages: The first issue is the so-called decellularization, which is best explored and resulted in a multitude of different protocols. The most promising approach seems to be the combination of pulsatile perfusion of the liver with Triton X-100 and SDS via hepatic artery and portal vein. Widely accepted parameters of quality control include the quantitative assessment of the DNA content and the visualization of eventually remaining nuclei confirmed by HE staining. Investigations regarding the composition of the extracellular matrix focused on histological determination of laminin, collagen, fibronectin and elastin and remained qualitatively. Repopulation is the second issue which is addressed. Selection of the most suitable cell type is a highly controversial topic. Currently, the highest potential is seen for progenitor and stem cells. Cells are infused into the scaffold and cultured under static conditions or in a bioreactor allowing dynamic perfusion of the scaffold. The quality of repopulation is mainly assessed by routine histology and basic functional assays. These promising results prompted to consider the use of a liver scaffold repopulated with human cells for pharmacological research. Transplantation of the (repopulated) scaffold is the third topic which is not yet widely addressed. Few studies report the heterotopic transplantation of repopulated liver tissue without vascular anastomosis. Even fewer studies deal with the heterotopic transplantation of a scaffold or a repopulated liver lobe. However, observation time was still limited to hours, and long-term graft survival has not been reported yet. These exciting results emphasize the potential of this new and promising strategy to create physiological models for pharmacological research and to generate liver grafts for the transplant community to treat organ failure. However, the scientific need for further development in the field of liver engineering is still tremendous.

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Section: The Decellularization Processmentioning

confidence: 99%

Section: The Decellularization Processmentioning

confidence: 99%

Section: Source Of Organs For Liver Engineeringmentioning

confidence: 99%

“…The time also ranged from less than 3 h [22] up to several days for total perfusion [27,31]. As the main route for the perfusion, the portal vein was used.…”

Section: The Decellularization Processmentioning

confidence: 99%

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Bioengineered Livers: A New Tool for Drug Testing and a Promising Solution to Meet the Growing Demand for Donor Organs

et al. 2016

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“…In all protocols, decellularization was achieved by perfusion of alkaline ionic or non-ionic detergents via the cannulated portal vein. To the best of our knowledge, we were the first group to report rat liver decellularization by selective perfusion via the portal vein and/or the rat hepatic artery 14 . Enabling the selective perfusion of the different vascular systems in the liver may enable better decellularization results and, furthermore, may play an important role in cellular repopulation.…”

Section: Introductionmentioning

confidence: 99%

Procedure for Decellularization of Rat Livers in an Oscillating-pressure Perfusion Device

Hillebrandt

Polenz

Butter

et al. 2015

JoVE

Self Cite

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Decellularization and recellularization of parenchymal organs may enable the generation of functional organs in vitro, and several protocols for rodent liver decellularization have already been published. We aimed to improve the decellularization process by construction of a proprietary perfusion device enabling selective perfusion via the portal vein and/or the hepatic artery. Furthermore, we sought to perform perfusion under oscillating surrounding pressure conditions to improve the homogeneity of decellularization. The homogeneity of perfusion decellularization has been an underestimated factor to date. During decellularization, areas within the organ that are poorly perfused may still contain cells, whereas the extracellular matrix (ECM) in well-perfused areas may already be affected by alkaline detergents. Oscillating pressure changes can mimic the intraabdominal pressure changes that occur during respiration to optimize microperfusion inside the liver. In the study presented here, decellularized rat liver matrices were analyzed by histological staining, DNA content analysis and corrosion casting. Perfusion via the hepatic artery showed more homogenous results than portal venous perfusion did. The application of oscillating pressure conditions improved the effectiveness of perfusion decellularization. Livers perfused via the hepatic artery and under oscillating pressure conditions showed the best results. The presented techniques for liver harvesting, cannulation and perfusion using our proprietary device enable sophisticated perfusion setups to improve decellularization and recellularization experiments in rat livers. Video LinkThe video component of this article can be found at

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Tissue and organ decellularization in regenerative medicine

Damodaran

Vermette

2018

Biotechnology Progress

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The advancement and improvement in decellularization methods can be attributed to the increasing demand for tissues and organs for transplantation. Decellularized tissues and organs, which are free of cells and genetic materials while retaining the complex ultrastructure of the extracellular matrix (ECM), can serve as scaffolds to subsequently embed cells for transplantation. They have the potential to mimic the native physiology of the targeted anatomic site. ECM from different tissues and organs harvested from various sources have been applied. Many techniques are currently involved in the decellularization process, which come along with their own advantages and disadvantages. This review focuses on recent developments in decellularization methods, the importance and nature of detergents used for decellularization, as well as on the role of the ECM either as merely a physical support or as a scaffold in retaining and providing cues for cell survival, differentiation and homeostasis. In addition, application, status, and perspectives on commercialization of bioproducts derived from decellularized tissues and organs are addressed. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:1494–1505, 2018

show abstract

Improved rat liver decellularization by arterial perfusion under oscillating pressure conditions

Cited by 57 publications

References 37 publications

Bioengineered Livers: A New Tool for Drug Testing and a Promising Solution to Meet the Growing Demand for Donor Organs

Bioengineered Livers: A New Tool for Drug Testing and a Promising Solution to Meet the Growing Demand for Donor Organs

Procedure for Decellularization of Rat Livers in an Oscillating-pressure Perfusion Device

Tissue and organ decellularization in regenerative medicine

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