Optimization of agitation speed in spinner flask for microcarrier structural integrity and expansion of induced pluripotent stem cells

Gupta, Poonam; Ismadi, Mohd Zulhilmi Paiz; Verma, Paul J.; Fouras, Andreas; Jadhav, Sameer; Bellare, Jayesh; Hourigan, Kerry

doi:10.1007/s10616-014-9750-z

Cited by 46 publications

(26 citation statements)

References 34 publications

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“…The characterization of commercial bioreactors has not been conducted until recent years [32][33][34]; furthermore, only limited mechanical parameters at a few speeds were investigated in these studies. Recently, Gupta et al [35] showed that the high rotational speeds could damage the microcarriers. The study also highlighted the optimum rotational speed corresponds to the highest mouse induced pluripotent cell yield.…”

Section: Introductionmentioning

confidence: 99%

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Experimental Characterisation of Fluid Mechanics in a Spinner Flask Bioreactor

2014

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The spinner flask bioreactor has been widely used in in vitro cell culturing processes due to its superiority in providing a homogeneous culture environment compared to traditional culturing methods. However, there is limited understanding of the flow fields in these bioreactors, and optimum culture conditions are yet to be determined. This article presents the experimental characterization of the flow field within a spinner flask at varying speeds (10 RPM to 80 RPM) and impeller positions. An optical, non-invasive measurement technique, Particle Image Velocimetry (PIV), was employed to illustrate the fluid flow and calculate the stresses and vorticity associated with the flow within the flask. The largest recirculation structure was observed in the meridional plane at the highest impeller position while the highest shear stress region was observed at the base of the spinner flask. The study provides an overview of the fluid structure within the spinner flask in the meridional and azimuthal planes. Furthermore, the results presented in this study give an accurate quantification of the range of stresses for the given impeller speeds. These results provide estimates of the biomechanical properties within the type of spinner flask used in many published cell studies.

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Section: Introductionmentioning

confidence: 99%

“…The study also highlighted the optimum rotational speed corresponds to the highest mouse induced pluripotent cell yield. Understanding the flow properties associated to the culture condition would enable researchers to gain advantage in optimising the culture process [35].…”

Section: Introductionmentioning

confidence: 99%

Experimental Characterisation of Fluid Mechanics in a Spinner Flask Bioreactor

2014

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“…The optimum concentration of MCs has not been reported, as it is dependent on cell type, MC properties, bioreactor type, and culture volume. However, a diverse range of ≈3000 MCs mL −1 to ≈15 000 MCs mL −1 is reported in the literature …”

Section: Concentration And/or Filtrationmentioning

confidence: 99%

Scaled‐Up Inertial Microfluidics: Retention System for Microcarrier‐Based Suspension Cultures

Moloudi

Yang

et al. 2019

Biotechnology Journal

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Recently, particle concentration and filtration using inertial microfluidics have drawn attention as an alternative to membrane and centrifugal technologies for industrial applications, where the target particle size varies between 1 µm and 500 µm. Inevitably, the bigger particle size (>50 µm) mandates scaling up the channel crosssection or hydraulic diameter (D H > 0.5 mm). The Dean-coupled inertial focusing dynamics in spiral microchannels is studied broadly; however, the impacts of secondary flow on particle migration in a scaled-up spiral channel is not fully elucidated. The mechanism of particle focusing inside scaled-up rectangular and trapezoidal spiral channels (i.e., 5-10× bigger than conventional microchannels) with an aim to develop a continuous and clog-free microfiltration system for bioprocessing is studied in detail. Herein, a unique focusing based on inflection point without the aid of sheath flow is reported. This new focusing mechanism, observed in the scaled-up channels, out-performs the conventional focusing scenarios in the previously reported trapezoidal and rectangular channels. Finally, as a proof-ofconcept, the utility of this device is showcased for the first time as a retention system for a cell-microcarrier (MC) suspension culture.

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“…More vigorous stirring and aeration are required to maintain the mass transfer rate of nutrients and gases for larger number of cells (Xing et al, 2009). However, this increases hydrodynamic shear stress, which produces adverse effects on cells, such as reduced proliferation, low viability, and uncontrolled differentiation of stem cells (Croughan, Hamel, & Wang, 1987;Gupta et al, 2016;Leung, Chen, Choo, Reuveny, & Oh, 2010;Ng, Berry, & Butler, 1996;O'Connor & Papoutsakis, 1992). The trade-off between the mass transfer rate and the hydrodynamic shear stress makes largescale expansion of shear-sensitive cells unreliable and leads to timeconsuming optimization of operating conditions on each expansion stage, as those factors are typically affected by the bioreactor's capacity.…”

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Hollow microcarriers for large‐scale expansion of anchorage‐dependent cells in a stirred bioreactor

YekrangSafakar

Acun

Choi

et al. 2018

Biotech & Bioengineering

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With recent advances in biotechnology, mammalian cells are used in biopharmaceutical industries to produce valuable protein therapeutics and investigated as effective therapeutic agents to permanently degenerative diseases in cell based therapy. In these exciting and actively expanding fields, a reliable, efficient, and affordable platform to culture mammalian cells on a large scale is one of the most vital necessities. To produce and maintain a very large population of anchorage-dependent cells, a microcarrier-based stirred tank bioreactor is commonly used. In this approach, the cells are exposed to harmful hydrodynamic shear stress in the bioreactor and the mass transfer rates of nutrients and gases in the bioreactor are often kept below an optimal level to prevent cellular damages from the shear stress. In this paper, a hollow microcarrier (HMC) is presented as a novel solution to protect cells from shear stress in stirred bioreactors, while ensuring sufficient and uniform mass transfer rate of gases and nutrients. HMC is a hollow microsphere and cells are cultured on its inner surface to be protected, while openings on the HMC provide sufficient exchange of media inside the HMC. As a proof of concept, we demonstrated the expansion of fibroblasts, NIH/3T3 and the expansion and cardiac differentiation of human induced pluripotent stem cells, along with detailed numerical analysis. We believe that the developed HMC can be a practical solution to enable large-scale expansion of shear-sensitive anchorage-dependent cells in an industrial scale with stirred bioreactors.

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Optimization of agitation speed in spinner flask for microcarrier structural integrity and expansion of induced pluripotent stem cells

Cited by 46 publications

References 34 publications

Experimental Characterisation of Fluid Mechanics in a Spinner Flask Bioreactor

Experimental Characterisation of Fluid Mechanics in a Spinner Flask Bioreactor

Scaled‐Up Inertial Microfluidics: Retention System for Microcarrier‐Based Suspension Cultures

Hollow microcarriers for large‐scale expansion of anchorage‐dependent cells in a stirred bioreactor

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