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Bioprinting Perfusion-Enabled Liver Equivalents for Advanced Organ-on-a-Chip Applications

Many tissue models have been developed to mimic liver-specific functions for metabolic and toxin conversion in in vitro assays. Most models represent a 2D environment rather than a complex 3D structure similar to native tissue. To overcome this issue, spheroid cultures have become the gold standard...

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Autores principales: Grix, Tobias, Ruppelt, Alicia, Thomas, Alexander, Amler, Anna-Klara, Noichl, Benjamin P., Lauster, Roland, Kloke, Lutz
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5924518/
https://www.ncbi.nlm.nih.gov/pubmed/29565814
http://dx.doi.org/10.3390/genes9040176
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author Grix, Tobias
Ruppelt, Alicia
Thomas, Alexander
Amler, Anna-Klara
Noichl, Benjamin P.
Lauster, Roland
Kloke, Lutz
author_facet Grix, Tobias
Ruppelt, Alicia
Thomas, Alexander
Amler, Anna-Klara
Noichl, Benjamin P.
Lauster, Roland
Kloke, Lutz
author_sort Grix, Tobias
collection PubMed
description Many tissue models have been developed to mimic liver-specific functions for metabolic and toxin conversion in in vitro assays. Most models represent a 2D environment rather than a complex 3D structure similar to native tissue. To overcome this issue, spheroid cultures have become the gold standard in tissue engineering. Unfortunately, spheroids are limited in size due to diffusion barriers in their dense structures, limiting nutrient and oxygen supply. Recent developments in bioprinting techniques have enabled us to engineer complex 3D structures with perfusion-enabled channel systems to ensure nutritional supply within larger, densely-populated tissue models. In this study, we present a proof-of-concept for the feasibility of bioprinting a liver organoid by combining HepaRG and human stellate cells in a stereolithographic printing approach, and show basic characterization under static cultivation conditions. Using standard tissue engineering analytics, such as immunohistology and qPCR, we found higher albumin and cytochrome P(450) 3A4 (CYP3A4) expression in bioprinted liver tissues compared to monolayer controls over a two-week cultivation period. In addition, the expression of tight junctions, liver-specific bile transporter multidrug resistance-associated protein 2 (MRP2), and overall metabolism (glucose, lactate, lactate dehydrogenase (LDH)) were found to be stable. Furthermore, we provide evidence for the perfusability of the organoids’ intrinsic channel system. These results motivate new approaches and further development in liver tissue engineering for advanced organ-on-a-chip applications and pharmaceutical developments.
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spelling pubmed-59245182018-05-03 Bioprinting Perfusion-Enabled Liver Equivalents for Advanced Organ-on-a-Chip Applications Grix, Tobias Ruppelt, Alicia Thomas, Alexander Amler, Anna-Klara Noichl, Benjamin P. Lauster, Roland Kloke, Lutz Genes (Basel) Article Many tissue models have been developed to mimic liver-specific functions for metabolic and toxin conversion in in vitro assays. Most models represent a 2D environment rather than a complex 3D structure similar to native tissue. To overcome this issue, spheroid cultures have become the gold standard in tissue engineering. Unfortunately, spheroids are limited in size due to diffusion barriers in their dense structures, limiting nutrient and oxygen supply. Recent developments in bioprinting techniques have enabled us to engineer complex 3D structures with perfusion-enabled channel systems to ensure nutritional supply within larger, densely-populated tissue models. In this study, we present a proof-of-concept for the feasibility of bioprinting a liver organoid by combining HepaRG and human stellate cells in a stereolithographic printing approach, and show basic characterization under static cultivation conditions. Using standard tissue engineering analytics, such as immunohistology and qPCR, we found higher albumin and cytochrome P(450) 3A4 (CYP3A4) expression in bioprinted liver tissues compared to monolayer controls over a two-week cultivation period. In addition, the expression of tight junctions, liver-specific bile transporter multidrug resistance-associated protein 2 (MRP2), and overall metabolism (glucose, lactate, lactate dehydrogenase (LDH)) were found to be stable. Furthermore, we provide evidence for the perfusability of the organoids’ intrinsic channel system. These results motivate new approaches and further development in liver tissue engineering for advanced organ-on-a-chip applications and pharmaceutical developments. MDPI 2018-03-22 /pmc/articles/PMC5924518/ /pubmed/29565814 http://dx.doi.org/10.3390/genes9040176 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Grix, Tobias
Ruppelt, Alicia
Thomas, Alexander
Amler, Anna-Klara
Noichl, Benjamin P.
Lauster, Roland
Kloke, Lutz
Bioprinting Perfusion-Enabled Liver Equivalents for Advanced Organ-on-a-Chip Applications
title Bioprinting Perfusion-Enabled Liver Equivalents for Advanced Organ-on-a-Chip Applications
title_full Bioprinting Perfusion-Enabled Liver Equivalents for Advanced Organ-on-a-Chip Applications
title_fullStr Bioprinting Perfusion-Enabled Liver Equivalents for Advanced Organ-on-a-Chip Applications
title_full_unstemmed Bioprinting Perfusion-Enabled Liver Equivalents for Advanced Organ-on-a-Chip Applications
title_short Bioprinting Perfusion-Enabled Liver Equivalents for Advanced Organ-on-a-Chip Applications
title_sort bioprinting perfusion-enabled liver equivalents for advanced organ-on-a-chip applications
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5924518/
https://www.ncbi.nlm.nih.gov/pubmed/29565814
http://dx.doi.org/10.3390/genes9040176
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