Design Criteria for Generating Physiologically Relevant In Vitro Models in Bioreactors
Abstract
:1. Introduction
1.1. In Vivo and In Vitro
1.2. Classifying the Microenvironment
Method | Description | Advantages | Disadvantages |
---|---|---|---|
Monolayer monocultures | Only one cell type plated in flat dishes | Cheap, standard | Loss of phenotype, absence of 3D architecture changes cell shape |
Tissue slice culture | Tissue slices are cultured in plates or on supports | In vivo environment is better preserved | Require hyperoxic conditions, viability limited in time |
Co-cultures | Monolayer cultures containing more than one cell type | Cell function is improved | Difficult to analyse the contribution of each cell type to co-culture function |
2. Mimicking Physiological Systems: The Rule of Ten
3. Scaling: Nano, Micro and Milli
Micro-Fluidics | Milli-Fluidics |
---|---|
Low shear by reduction of flow rate | Low shear, high flow rates |
Low nutrient turnover | High nutrient turnover |
High surface to volume ratio | Low surface to volume ratio |
Fiddly to assemble | Easy to assemble |
Presence of air bubbles | No air bubbles |
Low fluid volumes, saving on reagents | Higher volumes of media and reagents |
Easy quantification of cell products | Cell products may be harder to quantify |
4. Modelling 3D In Vitro Cell Cultures: Hepatocyte-Laden Hydrogels as a Reference
4.1. Modelled Configurations
4.2. Computational Mass Transport and Flow Model
4.2.1. Oxygen Transport and Consumption
Parameter | Symbol | Value | Units | References |
---|---|---|---|---|
Henry’s constant for oxygen | 1.32 × 10−3 | mol∙m−3∙mmHg−1 | [50] | |
Oxygen partial pressure in atmosphere | 159 | mmHg | [51] | |
Oxygen concentration in culture medium entering the system | 0.21 | mol/m3 | [31,32,52,53] | |
Oxygen diffusion in aqueous media | 3 × 10−9 | m2/s | [31,52,54] | |
Oxygen diffusion in the hydrogel construct | 1 × 10−9 | m2/s | [52,54,55,56,57,58] | |
Hepatocyte maximum oxygen consumption rate | Ω | 4.8 × 10−17 | mol∙cell−1∙s−1 | [49] |
Michaelis-Menten constant for oxygen consumption | Km | 7.39 × 10−3 | mol/m3 | [45,59,60] |
Critical oxygen concentration to account for cell necrosis | ccr | 2.64 × 10−3 | mol/m3 | [61] |
4.2.2. Fluid Dynamics
4.2.3. Model Implementation
Model | Surface | Boundary Condition |
---|---|---|
Oxygen convection and diffusion | System side walls | Insulation/symmetry (n · (−D∇c + cu) = 0) |
Interface between the hydrogel construct and the fluid sub-domain | Continuity | |
Fluid domain inlet | Constant oxygen concentration (c = 0.21 mol/m3) | |
Fluid domain outlet | Convective flux (n · (−D∇c) = 0) | |
Navier-Stokes | Solid-liquid interfaces | No slip (µ = 0) |
Fluid domain inlet | Normal inflow velocity (vin) | |
Fluid domain outlet | Pressure, no viscous stress (p0 = 0) |
4.3. CFD Model Refinement: Oxygen Diffusion through PDMS Walls
4.4. Results and Discussion
4.4.1. Theoretical Maximum Thickness for Functional Physiologically Relevant Hepatocyte Constructs
4.4.2. Fluid Dynamics in Micro- and Milli-Fluidic Systems
4.4.3. Oxygen Concentration Profiles in Micro- and Milli-Fluidic Systems
Modelled System | Micro-Fluidic | Milli-Fluidic | MCmB | |||
---|---|---|---|---|---|---|
OIW | OPW | OIW | OPW | OIW | OPW | |
Viable cells (%) | 2.6 | 100 | 17.8 | 78 | 60.5 | 85.4 |
Configuration | Flow Rate (m3/s) | Rin (mol/s) | Cell Number | Rcons (mol/s) | Rin/Rcons |
---|---|---|---|---|---|
Micro-fluidic | 2.84 × 10−14 | 1.42 × 10−13 | 5.60 × 104 | 4.48 × 10−13 | 0.3 |
Milli-fluidic | 6.40 × 10−13 | 3.20 × 10−12 | 5.60 × 104 | 4.48 × 10−13 | 7.1 |
MCmB | 3.00 × 10−9 | 1.50 × 10−8 | 2.47 × 106 | 1.98 × 10−11 | 758.3 |
5. The Allometric Approach
6. Conclusions
Acknowledgments
Author Contributions
Conflicts of Interest
References
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Mattei, G.; Giusti, S.; Ahluwalia, A. Design Criteria for Generating Physiologically Relevant In Vitro Models in Bioreactors. Processes 2014, 2, 548-569. https://doi.org/10.3390/pr2030548
Mattei G, Giusti S, Ahluwalia A. Design Criteria for Generating Physiologically Relevant In Vitro Models in Bioreactors. Processes. 2014; 2(3):548-569. https://doi.org/10.3390/pr2030548
Chicago/Turabian StyleMattei, Giorgio, Serena Giusti, and Arti Ahluwalia. 2014. "Design Criteria for Generating Physiologically Relevant In Vitro Models in Bioreactors" Processes 2, no. 3: 548-569. https://doi.org/10.3390/pr2030548