Cargando…

Fluid Dynamic Modeling to Support the Development of Flow-Based Hepatocyte Culture Systems for Metabolism Studies

Accurate prediction of metabolism is a significant outstanding challenge in toxicology. The best predictions are based on experimental data from in vitro systems using primary hepatocytes. The predictivity of the primary hepatocyte-based culture systems, however, is still limited due to well-known p...

Descripción completa

Detalles Bibliográficos
Autores principales: Pedersen, Jenny M., Shim, Yoo-Sik, Hans, Vaibhav, Phillips, Martin B., Macdonald, Jeffrey M., Walker, Glenn, Andersen, Melvin E., Clewell, Harvey J., Yoon, Miyoung
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5044513/
https://www.ncbi.nlm.nih.gov/pubmed/27747210
http://dx.doi.org/10.3389/fbioe.2016.00072
_version_ 1782456953460490240
author Pedersen, Jenny M.
Shim, Yoo-Sik
Hans, Vaibhav
Phillips, Martin B.
Macdonald, Jeffrey M.
Walker, Glenn
Andersen, Melvin E.
Clewell, Harvey J.
Yoon, Miyoung
author_facet Pedersen, Jenny M.
Shim, Yoo-Sik
Hans, Vaibhav
Phillips, Martin B.
Macdonald, Jeffrey M.
Walker, Glenn
Andersen, Melvin E.
Clewell, Harvey J.
Yoon, Miyoung
author_sort Pedersen, Jenny M.
collection PubMed
description Accurate prediction of metabolism is a significant outstanding challenge in toxicology. The best predictions are based on experimental data from in vitro systems using primary hepatocytes. The predictivity of the primary hepatocyte-based culture systems, however, is still limited due to well-known phenotypic instability and rapid decline of metabolic competence within a few hours. Dynamic flow bioreactors for three-dimensional cell cultures are thought to be better at recapitulating tissue microenvironments and show potential to improve in vivo extrapolations of chemical or drug toxicity based on in vitro test results. These more physiologically relevant culture systems hold potential for extending metabolic competence of primary hepatocyte cultures as well. In this investigation, we used computational fluid dynamics to determine the optimal design of a flow-based hepatocyte culture system for evaluating chemical metabolism in vitro. The main design goals were (1) minimization of shear stress experienced by the cells to maximize viability, (2) rapid establishment of a uniform distribution of test compound in the chamber, and (3) delivery of sufficient oxygen to cells to support aerobic respiration. Two commercially available flow devices – RealBio(®) and QuasiVivo(®) (QV) – and a custom developed fluidized bed bioreactor were simulated, and turbulence, flow characteristics, test compound distribution, oxygen distribution, and cellular oxygen consumption were analyzed. Experimental results from the bioreactors were used to validate the simulation results. Our results indicate that maintaining adequate oxygen supply is the most important factor to the long-term viability of liver bioreactor cultures. Cell density and system flow patterns were the major determinants of local oxygen concentrations. The experimental results closely corresponded to the in silico predictions. Of the three bioreactors examined in this study, we were able to optimize the experimental conditions for long-term hepatocyte cell culture using the QV bioreactor. This system facilitated the use of low system volumes coupled with higher flow rates. This design supports cellular respiration by increasing oxygen concentrations in the vicinity of the cells and facilitates long-term kinetic studies of low clearance test compounds. These two goals were achieved while simultaneously keeping the shear stress experienced by the cells within acceptable limits.
format Online
Article
Text
id pubmed-5044513
institution National Center for Biotechnology Information
language English
publishDate 2016
publisher Frontiers Media S.A.
record_format MEDLINE/PubMed
spelling pubmed-50445132016-10-14 Fluid Dynamic Modeling to Support the Development of Flow-Based Hepatocyte Culture Systems for Metabolism Studies Pedersen, Jenny M. Shim, Yoo-Sik Hans, Vaibhav Phillips, Martin B. Macdonald, Jeffrey M. Walker, Glenn Andersen, Melvin E. Clewell, Harvey J. Yoon, Miyoung Front Bioeng Biotechnol Bioengineering and Biotechnology Accurate prediction of metabolism is a significant outstanding challenge in toxicology. The best predictions are based on experimental data from in vitro systems using primary hepatocytes. The predictivity of the primary hepatocyte-based culture systems, however, is still limited due to well-known phenotypic instability and rapid decline of metabolic competence within a few hours. Dynamic flow bioreactors for three-dimensional cell cultures are thought to be better at recapitulating tissue microenvironments and show potential to improve in vivo extrapolations of chemical or drug toxicity based on in vitro test results. These more physiologically relevant culture systems hold potential for extending metabolic competence of primary hepatocyte cultures as well. In this investigation, we used computational fluid dynamics to determine the optimal design of a flow-based hepatocyte culture system for evaluating chemical metabolism in vitro. The main design goals were (1) minimization of shear stress experienced by the cells to maximize viability, (2) rapid establishment of a uniform distribution of test compound in the chamber, and (3) delivery of sufficient oxygen to cells to support aerobic respiration. Two commercially available flow devices – RealBio(®) and QuasiVivo(®) (QV) – and a custom developed fluidized bed bioreactor were simulated, and turbulence, flow characteristics, test compound distribution, oxygen distribution, and cellular oxygen consumption were analyzed. Experimental results from the bioreactors were used to validate the simulation results. Our results indicate that maintaining adequate oxygen supply is the most important factor to the long-term viability of liver bioreactor cultures. Cell density and system flow patterns were the major determinants of local oxygen concentrations. The experimental results closely corresponded to the in silico predictions. Of the three bioreactors examined in this study, we were able to optimize the experimental conditions for long-term hepatocyte cell culture using the QV bioreactor. This system facilitated the use of low system volumes coupled with higher flow rates. This design supports cellular respiration by increasing oxygen concentrations in the vicinity of the cells and facilitates long-term kinetic studies of low clearance test compounds. These two goals were achieved while simultaneously keeping the shear stress experienced by the cells within acceptable limits. Frontiers Media S.A. 2016-09-30 /pmc/articles/PMC5044513/ /pubmed/27747210 http://dx.doi.org/10.3389/fbioe.2016.00072 Text en Copyright © 2016 Pedersen, Shim, Hans, Phillips, Macdonald, Walker, Andersen, Clewell and Yoon. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Bioengineering and Biotechnology
Pedersen, Jenny M.
Shim, Yoo-Sik
Hans, Vaibhav
Phillips, Martin B.
Macdonald, Jeffrey M.
Walker, Glenn
Andersen, Melvin E.
Clewell, Harvey J.
Yoon, Miyoung
Fluid Dynamic Modeling to Support the Development of Flow-Based Hepatocyte Culture Systems for Metabolism Studies
title Fluid Dynamic Modeling to Support the Development of Flow-Based Hepatocyte Culture Systems for Metabolism Studies
title_full Fluid Dynamic Modeling to Support the Development of Flow-Based Hepatocyte Culture Systems for Metabolism Studies
title_fullStr Fluid Dynamic Modeling to Support the Development of Flow-Based Hepatocyte Culture Systems for Metabolism Studies
title_full_unstemmed Fluid Dynamic Modeling to Support the Development of Flow-Based Hepatocyte Culture Systems for Metabolism Studies
title_short Fluid Dynamic Modeling to Support the Development of Flow-Based Hepatocyte Culture Systems for Metabolism Studies
title_sort fluid dynamic modeling to support the development of flow-based hepatocyte culture systems for metabolism studies
topic Bioengineering and Biotechnology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5044513/
https://www.ncbi.nlm.nih.gov/pubmed/27747210
http://dx.doi.org/10.3389/fbioe.2016.00072
work_keys_str_mv AT pedersenjennym fluiddynamicmodelingtosupportthedevelopmentofflowbasedhepatocyteculturesystemsformetabolismstudies
AT shimyoosik fluiddynamicmodelingtosupportthedevelopmentofflowbasedhepatocyteculturesystemsformetabolismstudies
AT hansvaibhav fluiddynamicmodelingtosupportthedevelopmentofflowbasedhepatocyteculturesystemsformetabolismstudies
AT phillipsmartinb fluiddynamicmodelingtosupportthedevelopmentofflowbasedhepatocyteculturesystemsformetabolismstudies
AT macdonaldjeffreym fluiddynamicmodelingtosupportthedevelopmentofflowbasedhepatocyteculturesystemsformetabolismstudies
AT walkerglenn fluiddynamicmodelingtosupportthedevelopmentofflowbasedhepatocyteculturesystemsformetabolismstudies
AT andersenmelvine fluiddynamicmodelingtosupportthedevelopmentofflowbasedhepatocyteculturesystemsformetabolismstudies
AT clewellharveyj fluiddynamicmodelingtosupportthedevelopmentofflowbasedhepatocyteculturesystemsformetabolismstudies
AT yoonmiyoung fluiddynamicmodelingtosupportthedevelopmentofflowbasedhepatocyteculturesystemsformetabolismstudies