Characterization of the Aeration and Hydrodynamics in Vertical-Wheel™ Bioreactors

In this work, the oxygen transport and hydrodynamic flow of the PBS Vertical-Wheel MINI™ 0.1 bioreactor were characterized using experimental data and computational fluid dynamics simulations. Data acquired from spectroscopy-based oxygenation measurements was compared with data obtained from 3D simulations with a rigid-lid approximation and LES-WALE turbulence modeling, using the open-source software OpenFOAM-8. The mass transfer coefficients were determined for a range of stirring speeds between 10 and 100 rpm and for working volumes between 60 and 100 mL. Additionally, boundary condition, mesh refinement, and temperature variation studies were performed. Lastly, cell size, energy dissipation rate, and shear stress fields were calculated to determine optimal hydrodynamic conditions for culture. The experimental results demonstrate that the kL can be predicted using Sh=1.68Re0.551Sc13G1.18, with a mean absolute error of 2.08%. Using the simulations and a correction factor of 0.473, the expression can be correlated to provide equally valid results. To directly obtain them from simulations, a partial slip boundary condition can be tuned, ensuring better near-surface velocity profiles or, alternatively, by deeply refining the mesh. Temperature variation studies support the use of this correlation for temperatures up to 37 °C by using a Schmidt exponent of 1/3. Finally, the flow was characterized as transitional with diverse mixing mechanisms that ensure homogeneity and suspension quality, and the results obtained are in agreement with previous studies that employed RANS models. Overall, this work provides new data regarding oxygen mass transfer and hydrodynamics in the Vertical-Wheel bioreactor, as well as new insights for air-water mass transfer modeling in systems with low interface deformation, and a computational model that can be used for further studies.

[1]  N. Muhammad Finite volume method for simulation of flowing fluid via OpenFOAM , 2021, The European Physical Journal Plus.

[2]  Tiffany Dang,et al.  Author response for "Computational fluid dynamic characterization of vertical‐wheel bioreactors used for effective scale‐up of human induced pluripotent stem cell aggregate culture" , 2021 .

[3]  N. Bernardes,et al.  Scalable Production of Human Mesenchymal Stromal Cell-Derived Extracellular Vesicles Under Serum-/Xeno-Free Conditions in a Microcarrier-Based Bioreactor Culture System , 2020, Frontiers in Cell and Developmental Biology.

[4]  Breanna S Borys,et al.  Overcoming bioprocess bottlenecks in the large-scale expansion of high-quality hiPSC aggregates in vertical-wheel stirred suspension bioreactors , 2020, Stem cell research & therapy.

[5]  F. Ferreira,et al.  Scalable Manufacturing of Human Mesenchymal Stromal Cells in the Vertical-Wheel Bioreactor System: An Experimental and Economic Approach. , 2020, Biotechnology journal.

[6]  H. Tsutsui,et al.  Hydrodynamic characterization within a spinner flask and a rotary wall vessel for stem cell culture , 2020, Biochemical Engineering Journal.

[7]  Breanna S Borys,et al.  Using Computational Fluid Dynamics (CFD) Modeling to understand Murine Embryonic Stem Cell Aggregate Size and Pluripotency Distributions in Stirred Suspension Bioreactors. , 2019, Journal of biotechnology.

[8]  Brian Lee,et al.  Strategies for the expansion of human induced pluripotent stem cells as aggregates in single-use Vertical-Wheel™ bioreactors , 2019, Journal of Biological Engineering.

[9]  Joaquim M. S. Cabral,et al.  Scalable culture of human induced pluripotent cells on microcarriers under xeno-free conditions using single-use vertical-wheel™ bioreactors , 2018, Journal of Chemical Technology & Biotechnology.

[10]  F. Bombardelli,et al.  On the Values for the Turbulent Schmidt Number in Environmental Flows , 2017 .

[11]  J. Park,et al.  Usage of Human Mesenchymal Stem Cells in Cell-based Therapy: Advantages and Disadvantages , 2017, Development & reproduction.

[12]  J. D. Berry,et al.  Characterisation of stresses on microcarriers in a stirred bioreactor , 2016 .

[13]  An Le,et al.  Computational Fluid Dynamics Modeling of Scalable Stirred Suspension Bioreactors for Pluripotent Stem Cell Expansion , 2016 .

[14]  L. Quek,et al.  Metabolic Profiling and Flux Analysis of MEL-2 Human Embryonic Stem Cells during Exponential Growth at Physiological and Atmospheric Oxygen Concentrations , 2014, PloS one.

[15]  Jincheng Wu,et al.  Oxygen Transport and Stem Cell Aggregation in Stirred-Suspension Bioreactor Cultures , 2014, PloS one.

[16]  Diego Donzis,et al.  The Turbulent Schmidt Number , 2014 .

[17]  M. S. Kallos,et al.  Shear stress influences the pluripotency of murine embryonic stem cells in stirred suspension bioreactors , 2014, Journal of tissue engineering and regenerative medicine.

[18]  Hyun Dong Kim,et al.  Measurement of dissolved oxygen diffusion coefficient in a microchannel using UV-LED induced fluorescence method , 2013 .

[19]  M. Teitell,et al.  Measuring energy metabolism in cultured cells, including human pluripotent stem cells and differentiated cells , 2012, Nature Protocols.

[20]  Michael S Kallos,et al.  Mass Transfer Limitations in Embryoid Bodies during Human Embryonic Stem Cell Differentiation , 2012, Cells Tissues Organs.

[21]  G. Schatten,et al.  Energy Metabolism in Human Pluripotent Stem Cells and Their Differentiated Counterparts , 2011, PloS one.

[22]  Gordon A. Hill,et al.  Modelling Oxygen Transfer and Aerobic Growth in Shake Flasks and Well‐Mixed Bioreactors , 2008 .

[23]  Hu Zhang,et al.  Computational‐fluid‐dynamics (CFD) analysis of mixing and gas–liquid mass transfer in shake flasks , 2005, Biotechnology and applied biochemistry.

[24]  G. Rao,et al.  A study of oxygen transfer in shake flasks using a non‐invasive oxygen sensor , 2003, Biotechnology and bioengineering.

[25]  Andrew T. Jessup,et al.  Microscale wave breaking and air‐water gas transfer , 2001 .

[26]  S. Middleman,et al.  An Introduction to Mass and Heat Transfer: Principles of Analysis and Design , 1997 .

[27]  L. Ju,et al.  Oxygen diffusion coefficient and solubility in n-hexadecane. , 1989, Biotechnology and bioengineering.

[28]  P. Liss,et al.  Models for air-water gas transfer: an experimental investigation , 1984 .

[29]  J. Kestin,et al.  Viscosity of Liquid Water in the Range - 8 C to 150 C, , 1978 .

[30]  C. St-Denis,et al.  Diffusivity of oxygen in water , 1971 .

[31]  D. Ratkowsky,et al.  Effect of transverse vibration upon the rate of mass transfer from horizontal cylinders , 1968 .

[32]  J. Davies,et al.  The effects of surface films in damping eddies at a free surface of a turbulent liquid , 1966, Proceedings of the Royal Society of London. Series A. Mathematical and Physical Sciences.

[33]  P. Ziherl,et al.  Transport Phenomena , 2019, Solved Problems in Thermodynamics and Statistical Physics.

[34]  P. Liovic,et al.  FLUID FLOW AND STRESSES ON MICROCARRIERS IN SPINNER FLASK BIOREACTORS , 2012 .

[35]  K. McCloskey,et al.  Can shear stress direct stem cell fate? , 2009, Biotechnology progress.

[36]  Robert S. Cherry,et al.  Hydrodynamic effects on cells in agitated tissue culture reactors , 1986 .

[37]  T. G. Theofanous,et al.  Conceptual Models of Gas Exchange , 1984 .

[38]  Robert E. Wilson,et al.  Fundamentals of momentum, heat, and mass transfer , 1969 .