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Cell-Type-Specific Quantification of a Scaffold-Based 3D Liver Co-Culture

In order to increase the metabolic activity of human hepatocytes and liver cancer cell lines, many approaches have been reported in recent years. The metabolic activity could be increased mainly by cultivating the cells in 3D systems or co-cultures (with other cell lines). However, if the system bec...

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Autores principales: Ruoß, Marc, Kieber, Vanessa, Rebholz, Silas, Linnemann, Caren, Rinderknecht, Helen, Häussling, Victor, Häcker, Marina, Olde Damink, Leon H. H., Ehnert, Sabrina, Nussler, Andreas K.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2019
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7189675/
https://www.ncbi.nlm.nih.gov/pubmed/31878071
http://dx.doi.org/10.3390/mps3010001
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author Ruoß, Marc
Kieber, Vanessa
Rebholz, Silas
Linnemann, Caren
Rinderknecht, Helen
Häussling, Victor
Häcker, Marina
Olde Damink, Leon H. H.
Ehnert, Sabrina
Nussler, Andreas K.
author_facet Ruoß, Marc
Kieber, Vanessa
Rebholz, Silas
Linnemann, Caren
Rinderknecht, Helen
Häussling, Victor
Häcker, Marina
Olde Damink, Leon H. H.
Ehnert, Sabrina
Nussler, Andreas K.
author_sort Ruoß, Marc
collection PubMed
description In order to increase the metabolic activity of human hepatocytes and liver cancer cell lines, many approaches have been reported in recent years. The metabolic activity could be increased mainly by cultivating the cells in 3D systems or co-cultures (with other cell lines). However, if the system becomes more complex, it gets more difficult to quantify the number of cells (e.g., on a 3D matrix). Until now, it has been impossible to quantify different cell types individually in 3D co-culture systems. Therefore, we developed a PCR-based method that allows the quantification of HepG2 cells and 3T3-J2 cells separately in a 3D scaffold culture. Moreover, our results show that this method allows better comparability between 2D and 3D cultures in comparison to the often-used approaches based on metabolic activity measurements, such as the conversion of resazurin.
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spelling pubmed-71896752020-05-01 Cell-Type-Specific Quantification of a Scaffold-Based 3D Liver Co-Culture Ruoß, Marc Kieber, Vanessa Rebholz, Silas Linnemann, Caren Rinderknecht, Helen Häussling, Victor Häcker, Marina Olde Damink, Leon H. H. Ehnert, Sabrina Nussler, Andreas K. Methods Protoc Article In order to increase the metabolic activity of human hepatocytes and liver cancer cell lines, many approaches have been reported in recent years. The metabolic activity could be increased mainly by cultivating the cells in 3D systems or co-cultures (with other cell lines). However, if the system becomes more complex, it gets more difficult to quantify the number of cells (e.g., on a 3D matrix). Until now, it has been impossible to quantify different cell types individually in 3D co-culture systems. Therefore, we developed a PCR-based method that allows the quantification of HepG2 cells and 3T3-J2 cells separately in a 3D scaffold culture. Moreover, our results show that this method allows better comparability between 2D and 3D cultures in comparison to the often-used approaches based on metabolic activity measurements, such as the conversion of resazurin. MDPI 2019-12-23 /pmc/articles/PMC7189675/ /pubmed/31878071 http://dx.doi.org/10.3390/mps3010001 Text en © 2019 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
Ruoß, Marc
Kieber, Vanessa
Rebholz, Silas
Linnemann, Caren
Rinderknecht, Helen
Häussling, Victor
Häcker, Marina
Olde Damink, Leon H. H.
Ehnert, Sabrina
Nussler, Andreas K.
Cell-Type-Specific Quantification of a Scaffold-Based 3D Liver Co-Culture
title Cell-Type-Specific Quantification of a Scaffold-Based 3D Liver Co-Culture
title_full Cell-Type-Specific Quantification of a Scaffold-Based 3D Liver Co-Culture
title_fullStr Cell-Type-Specific Quantification of a Scaffold-Based 3D Liver Co-Culture
title_full_unstemmed Cell-Type-Specific Quantification of a Scaffold-Based 3D Liver Co-Culture
title_short Cell-Type-Specific Quantification of a Scaffold-Based 3D Liver Co-Culture
title_sort cell-type-specific quantification of a scaffold-based 3d liver co-culture
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7189675/
https://www.ncbi.nlm.nih.gov/pubmed/31878071
http://dx.doi.org/10.3390/mps3010001
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