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A Perfusion Bioreactor System for Cell Seeding and Oxygen-Controlled Cultivation of Three-Dimensional Cell Cultures

Bioreactor systems facilitate three-dimensional (3D) cell culture by coping with limitations of static cultivation techniques. To allow for the investigation of proper cultivation conditions and the reproducible generation of tissue-engineered grafts, a bioreactor system, which comprises the control...

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Autores principales: Schmid, Jakob, Schwarz, Sascha, Meier-Staude, Robert, Sudhop, Stefanie, Clausen-Schaumann, Hauke, Schieker, Matthias, Huber, Robert
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Mary Ann Liebert, Inc., publishers 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6208160/
https://www.ncbi.nlm.nih.gov/pubmed/30234443
http://dx.doi.org/10.1089/ten.tec.2018.0204
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author Schmid, Jakob
Schwarz, Sascha
Meier-Staude, Robert
Sudhop, Stefanie
Clausen-Schaumann, Hauke
Schieker, Matthias
Huber, Robert
author_facet Schmid, Jakob
Schwarz, Sascha
Meier-Staude, Robert
Sudhop, Stefanie
Clausen-Schaumann, Hauke
Schieker, Matthias
Huber, Robert
author_sort Schmid, Jakob
collection PubMed
description Bioreactor systems facilitate three-dimensional (3D) cell culture by coping with limitations of static cultivation techniques. To allow for the investigation of proper cultivation conditions and the reproducible generation of tissue-engineered grafts, a bioreactor system, which comprises the control of crucial cultivation parameters in independent-operating parallel bioreactors, is beneficial. Furthermore, the use of a bioreactor as an automated cell seeding tool enables even cell distributions on stable scaffolds. In this study, we developed a perfusion microbioreactor system, which enables the cultivation of 3D cell cultures in an oxygen-controlled environment in up to four independent-operating bioreactors. Therefore, perfusion microbioreactors were designed with the help of computer-aided design, and manufactured using the 3D printing technologies stereolithography and fused deposition modeling. A uniform flow distribution in the microbioreactor was shown using a computational fluid dynamics model. For oxygen measurements, microsensors were integrated in the bioreactors to measure the oxygen concentration (OC) in the geometric center of the 3D cell cultures. To control the OC in each bioreactor independently, an automated feedback loop was developed, which adjusts the perfusion velocity according to the oxygen sensor signal. Furthermore, an automated cell seeding protocol was implemented to facilitate the even distribution of cells within a stable scaffold in a reproducible way. As proof of concept, the human mesenchymal stem cell line SCP-1 was seeded on bovine cancellous bone matrix of 1 cm(3) and cultivated in the developed microbioreactor system at different oxygen levels. The oxygen control was capable to maintain preset oxygen levels ±0.5% over a cultivation period of several days. Using the automated cell seeding procedure resulted in evenly distributed cells within a stable scaffold. In summary, the developed microbioreactor system enables the cultivation of 3D cell cultures in an automated and thus reproducible way by providing up to four independently operating, oxygen-controlled bioreactors. In combination with the automated cell seeding procedure, the bioreactor system opens up new possibilities to conduct more reproducible experiments to investigate optimal cultivation parameters and to generate tissue-engineering grafts in an oxygen-controlled environment.
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spelling pubmed-62081602018-10-31 A Perfusion Bioreactor System for Cell Seeding and Oxygen-Controlled Cultivation of Three-Dimensional Cell Cultures Schmid, Jakob Schwarz, Sascha Meier-Staude, Robert Sudhop, Stefanie Clausen-Schaumann, Hauke Schieker, Matthias Huber, Robert Tissue Eng Part C Methods Methods Articles Bioreactor systems facilitate three-dimensional (3D) cell culture by coping with limitations of static cultivation techniques. To allow for the investigation of proper cultivation conditions and the reproducible generation of tissue-engineered grafts, a bioreactor system, which comprises the control of crucial cultivation parameters in independent-operating parallel bioreactors, is beneficial. Furthermore, the use of a bioreactor as an automated cell seeding tool enables even cell distributions on stable scaffolds. In this study, we developed a perfusion microbioreactor system, which enables the cultivation of 3D cell cultures in an oxygen-controlled environment in up to four independent-operating bioreactors. Therefore, perfusion microbioreactors were designed with the help of computer-aided design, and manufactured using the 3D printing technologies stereolithography and fused deposition modeling. A uniform flow distribution in the microbioreactor was shown using a computational fluid dynamics model. For oxygen measurements, microsensors were integrated in the bioreactors to measure the oxygen concentration (OC) in the geometric center of the 3D cell cultures. To control the OC in each bioreactor independently, an automated feedback loop was developed, which adjusts the perfusion velocity according to the oxygen sensor signal. Furthermore, an automated cell seeding protocol was implemented to facilitate the even distribution of cells within a stable scaffold in a reproducible way. As proof of concept, the human mesenchymal stem cell line SCP-1 was seeded on bovine cancellous bone matrix of 1 cm(3) and cultivated in the developed microbioreactor system at different oxygen levels. The oxygen control was capable to maintain preset oxygen levels ±0.5% over a cultivation period of several days. Using the automated cell seeding procedure resulted in evenly distributed cells within a stable scaffold. In summary, the developed microbioreactor system enables the cultivation of 3D cell cultures in an automated and thus reproducible way by providing up to four independently operating, oxygen-controlled bioreactors. In combination with the automated cell seeding procedure, the bioreactor system opens up new possibilities to conduct more reproducible experiments to investigate optimal cultivation parameters and to generate tissue-engineering grafts in an oxygen-controlled environment. Mary Ann Liebert, Inc., publishers 2018-10-01 2018-10-17 /pmc/articles/PMC6208160/ /pubmed/30234443 http://dx.doi.org/10.1089/ten.tec.2018.0204 Text en © Jakob Schmid et al. 2018; Published by Mary Ann Liebert, Inc. This Open Access article is distributed under the terms of the Creative Commons License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methods Articles
Schmid, Jakob
Schwarz, Sascha
Meier-Staude, Robert
Sudhop, Stefanie
Clausen-Schaumann, Hauke
Schieker, Matthias
Huber, Robert
A Perfusion Bioreactor System for Cell Seeding and Oxygen-Controlled Cultivation of Three-Dimensional Cell Cultures
title A Perfusion Bioreactor System for Cell Seeding and Oxygen-Controlled Cultivation of Three-Dimensional Cell Cultures
title_full A Perfusion Bioreactor System for Cell Seeding and Oxygen-Controlled Cultivation of Three-Dimensional Cell Cultures
title_fullStr A Perfusion Bioreactor System for Cell Seeding and Oxygen-Controlled Cultivation of Three-Dimensional Cell Cultures
title_full_unstemmed A Perfusion Bioreactor System for Cell Seeding and Oxygen-Controlled Cultivation of Three-Dimensional Cell Cultures
title_short A Perfusion Bioreactor System for Cell Seeding and Oxygen-Controlled Cultivation of Three-Dimensional Cell Cultures
title_sort perfusion bioreactor system for cell seeding and oxygen-controlled cultivation of three-dimensional cell cultures
topic Methods Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6208160/
https://www.ncbi.nlm.nih.gov/pubmed/30234443
http://dx.doi.org/10.1089/ten.tec.2018.0204
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