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A Perfusion Bioreactor for Longitudinal Monitoring of Bioengineered Liver Constructs

In the field of in vitro liver disease models, decellularised organ scaffolds maintain the original biomechanical and biological properties of the extracellular matrix and are established supports for in vitro cell culture. However, tissue engineering approaches based on whole organ decellularized s...

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Autores principales: Sassi, Lisa, Ajayi, Omolola, Campinoti, Sara, Natarajan, Dipa, McQuitty, Claire, Siena, Riccardo Rayan, Mantero, Sara, De Coppi, Paolo, Pellegata, Alessandro F., Chokshi, Shilpa, Urbani, Luca
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7912543/
https://www.ncbi.nlm.nih.gov/pubmed/33494337
http://dx.doi.org/10.3390/nano11020275
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author Sassi, Lisa
Ajayi, Omolola
Campinoti, Sara
Natarajan, Dipa
McQuitty, Claire
Siena, Riccardo Rayan
Mantero, Sara
De Coppi, Paolo
Pellegata, Alessandro F.
Chokshi, Shilpa
Urbani, Luca
author_facet Sassi, Lisa
Ajayi, Omolola
Campinoti, Sara
Natarajan, Dipa
McQuitty, Claire
Siena, Riccardo Rayan
Mantero, Sara
De Coppi, Paolo
Pellegata, Alessandro F.
Chokshi, Shilpa
Urbani, Luca
author_sort Sassi, Lisa
collection PubMed
description In the field of in vitro liver disease models, decellularised organ scaffolds maintain the original biomechanical and biological properties of the extracellular matrix and are established supports for in vitro cell culture. However, tissue engineering approaches based on whole organ decellularized scaffolds are hampered by the scarcity of appropriate bioreactors that provide controlled 3D culture conditions. Novel specific bioreactors are needed to support long-term culture of bioengineered constructs allowing non-invasive longitudinal monitoring. Here, we designed and validated a specific bioreactor for long-term 3D culture of whole liver constructs. Whole liver scaffolds were generated by perfusion decellularisation of rat livers. Scaffolds were seeded with Luc(+)HepG2 and primary human hepatocytes and cultured in static or dynamic conditions using the custom-made bioreactor. The bioreactor included a syringe pump, for continuous unidirectional flow, and a circuit built to allow non-invasive monitoring of culture parameters and media sampling. The bioreactor allowed non-invasive analysis of cell viability, distribution, and function of Luc(+)HepG2-bioengineered livers cultured for up to 11 days. Constructs cultured in dynamic conditions in the bioreactor showed significantly higher cell viability, measured with bioluminescence, distribution, and functionality (determined by albumin production and expression of CYP enzymes) in comparison to static culture conditions. Finally, our bioreactor supports primary human hepatocyte viability and function for up to 30 days, when seeded in the whole liver scaffolds. Overall, our novel bioreactor is capable of supporting cell survival and metabolism and is suitable for liver tissue engineering for the development of 3D liver disease models.
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spelling pubmed-79125432021-02-28 A Perfusion Bioreactor for Longitudinal Monitoring of Bioengineered Liver Constructs Sassi, Lisa Ajayi, Omolola Campinoti, Sara Natarajan, Dipa McQuitty, Claire Siena, Riccardo Rayan Mantero, Sara De Coppi, Paolo Pellegata, Alessandro F. Chokshi, Shilpa Urbani, Luca Nanomaterials (Basel) Article In the field of in vitro liver disease models, decellularised organ scaffolds maintain the original biomechanical and biological properties of the extracellular matrix and are established supports for in vitro cell culture. However, tissue engineering approaches based on whole organ decellularized scaffolds are hampered by the scarcity of appropriate bioreactors that provide controlled 3D culture conditions. Novel specific bioreactors are needed to support long-term culture of bioengineered constructs allowing non-invasive longitudinal monitoring. Here, we designed and validated a specific bioreactor for long-term 3D culture of whole liver constructs. Whole liver scaffolds were generated by perfusion decellularisation of rat livers. Scaffolds were seeded with Luc(+)HepG2 and primary human hepatocytes and cultured in static or dynamic conditions using the custom-made bioreactor. The bioreactor included a syringe pump, for continuous unidirectional flow, and a circuit built to allow non-invasive monitoring of culture parameters and media sampling. The bioreactor allowed non-invasive analysis of cell viability, distribution, and function of Luc(+)HepG2-bioengineered livers cultured for up to 11 days. Constructs cultured in dynamic conditions in the bioreactor showed significantly higher cell viability, measured with bioluminescence, distribution, and functionality (determined by albumin production and expression of CYP enzymes) in comparison to static culture conditions. Finally, our bioreactor supports primary human hepatocyte viability and function for up to 30 days, when seeded in the whole liver scaffolds. Overall, our novel bioreactor is capable of supporting cell survival and metabolism and is suitable for liver tissue engineering for the development of 3D liver disease models. MDPI 2021-01-21 /pmc/articles/PMC7912543/ /pubmed/33494337 http://dx.doi.org/10.3390/nano11020275 Text en © 2021 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
Sassi, Lisa
Ajayi, Omolola
Campinoti, Sara
Natarajan, Dipa
McQuitty, Claire
Siena, Riccardo Rayan
Mantero, Sara
De Coppi, Paolo
Pellegata, Alessandro F.
Chokshi, Shilpa
Urbani, Luca
A Perfusion Bioreactor for Longitudinal Monitoring of Bioengineered Liver Constructs
title A Perfusion Bioreactor for Longitudinal Monitoring of Bioengineered Liver Constructs
title_full A Perfusion Bioreactor for Longitudinal Monitoring of Bioengineered Liver Constructs
title_fullStr A Perfusion Bioreactor for Longitudinal Monitoring of Bioengineered Liver Constructs
title_full_unstemmed A Perfusion Bioreactor for Longitudinal Monitoring of Bioengineered Liver Constructs
title_short A Perfusion Bioreactor for Longitudinal Monitoring of Bioengineered Liver Constructs
title_sort perfusion bioreactor for longitudinal monitoring of bioengineered liver constructs
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7912543/
https://www.ncbi.nlm.nih.gov/pubmed/33494337
http://dx.doi.org/10.3390/nano11020275
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