Computational fluid model incorporating liver metabolic activities in perfusion bioreactor

Hsu, Myat Noe; Tan, Guo-Dong Sean; Tania, Marshella; Birgersson, Erik; Leo, Hwa Liang

doi:10.1002/bit.25157

Cited by 16 publications

(18 citation statements)

References 33 publications

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“…23 Considering the increase of albumin production within days 12 to 15 as well as the good viability of the cells within at least two weeks of cultivation, we suggest that the cells do not suffer from oxygen or nutrient deprivation. In the literature, albumin secretion rates for hepatocytes cultivated in biochips range from 3.6 pg/cell/day (HepG2 cells) over 2.6-19 pg/cell/day (primary human hepatocytes) to 10-60 pg/cell/day (primary rat hepatocytes), and T€ or€ ok et al estimated the in vivo secretion rate to 17.8 pg/cell/day.…”

Section: Discussionmentioning

confidence: 78%

On-chip three-dimensional cell culture in phaseguides improves hepatocyte functions in vitro

et al. 2015

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The in vitro study of liver functions and liver cell specific responses to external stimuli deals with the problem to preserve the in vivo functions of primary hepatocytes. In this study, we used the biochip OrganoPlate TM (MIMETAS) that combines different advantages for the cultivation of hepatocytes in vitro: (1) the perfusion flow is achieved without a pump allowing easy handling and placement in the incubator; (2) the phaseguides allow plating of matrix-embedded cells in lanes adjacent to the perfusion flow without physical barrier; and (3) the matrix-embedding ensures indirect contact of the cells to the flow. In order to evaluate the applicability of this biochip for the study of hepatocyte's functions, Matrigel TM -embedded HepG2 cells were cultured over three weeks in this biochip and compared to a static Matrigel culture (3D) and a monolayer culture (2D). Chip-cultured cells grew in spheroid-like structures and were characterized by the formation of bile canaliculi and a high viability over 14 days. Hepatocyte-specific physiology was achieved as determined by an increase in albumin production. Improved detoxification metabolism was demonstrated by strongly increased cytochrome P450 activity and urea production. Additionally, chip-cultured cells displayed increased sensitivity to acetaminophen. Altogether, the OrganoPlate seems to be a very useful alternative for the cultivation of hepatocytes, as their behavior was strongly improved over 2D and static 3D cultures and the results were largely comparable and partly superior to the previous reports on biochip-cultured hepatocytes. As for the low technical needs, this platform has the appearance of being highly applicable for further studies of hepatocytes' responses to external stimuli. V C 2015 AIP Publishing LLC. [http://dx

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Section: Discussionmentioning

confidence: 78%

On-chip three-dimensional cell culture in phaseguides improves hepatocyte functions in vitro

et al. 2015

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“…Many of the bioreactor systems are flow-based culture systems, and computational fluid dynamics (CFD) modeling is well suited to optimize bioreactor design as it provides a tool to describe fluid dynamics in the engineered culture devices (Kang et al, 2013 ; Hsu et al, 2014 ). CFD modeling allows parameter values to be varied and the effects on the system to be characterized rapidly.…”

Section: Introductionmentioning

confidence: 99%

“…It can also be used to describe spatiotemporal distributions of a test compound or oxygen within the system. CFD and other computational methods have previously been applied to enhance our understanding of system parameters fundamental for cell responses within complex 3D culture environments (Kang et al, 2013 ; Hsu et al, 2014 ).…”

Section: Introductionmentioning

confidence: 99%

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

Pedersen

Shim

Hans

et al. 2016

Front. Bioeng. Biotechnol.

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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.

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“…First, we consider a reaction governed by nonlinear saturable binding kinetics, suitable for describing the effect of a wide range of drugs whose mode of action is governed by ligand-receptor interactions. Then, we consider a reaction governed by M-M kinetics, commonly used in the literature to describe O 2 consumption [5,7,8,[12][13][14] and paracetamol (APAP) metabolism [16,17].…”

Section: Solute Reaction With the Cellsmentioning

confidence: 99%

Mathematical modelling of fluid flow and solute transport to define operating parameters forin vitroperfusion cell culture systems

et al. 2020

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In recent years, there has been a move away from the use of static in vitro two-dimensional cell culture models for testing the chemical safety and efficacy of drugs. Such models are increasingly being replaced by more physiologically relevant cell culture systems featuring dynamic flow and/or three-dimensional structures of cells. While it is acknowledged that such systems provide a more realistic environment within which to test drugs, progress is being hindered by a lack of understanding of the physical and chemical environment that the cells are exposed to. Mathematical and computational modelling may be exploited in this regard to unravel the dependency of the cell response on spatio-temporal differences in chemical and mechanical cues, thereby assisting with the understanding and design of these systems. In this paper, we present a mathematical modelling framework that characterizes the fluid flow and solute transport in perfusion bioreactors featuring an inlet and an outlet. To demonstrate the utility of our model, we simulated the fluid dynamics and solute concentration profiles for a variety of different flow rates, inlet solute concentrations and cell types within a specific commercial bioreactor chamber. Our subsequent analysis has elucidated the basic relationship between inlet flow rate and cell surface flow speed, shear stress and solute concentrations, allowing us to derive simple but useful relationships that enable prediction of the behaviour of the system under a variety of experimental conditions, prior to experimentation. We describe how the model may used by experimentalists to define operating parameters for their particular perfusion cell culture systems and highlight some operating conditions that should be avoided. Finally, we critically comment on the limitations of mathematical and computational modelling in this field, and the challenges associated with the adoption of such methods.

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Computational fluid model incorporating liver metabolic activities in perfusion bioreactor

Cited by 16 publications

References 33 publications

On-chip three-dimensional cell culture in phaseguides improves hepatocyte functions in vitro

On-chip three-dimensional cell culture in phaseguides improves hepatocyte functions in vitro

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

Mathematical modelling of fluid flow and solute transport to define operating parameters forin vitroperfusion cell culture systems

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