Multiphase mixing characteristics in a microcarrier-based stirred tank bioreactor suitable for human mesenchymal stem cell expansion

Grein, Tanja A.; Leber, Jasmin; Blumenstock, Miriam; Petry, Florian; Weidner, Tobias; Salzig, Denise; Czermak, Peter

doi:10.1016/j.procbio.2016.05.010

Cited by 28 publications

(18 citation statements)

References 37 publications

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“…These have a controlled power input, ensuring the homogeneous mixing of suspension cells and anchorage-dependent adherent cells growing on microcarriers. However, dynamic cultivation systems are a double-edged sword because the homogeneity of cell growth comes at the expense of high shear stress [56,57]. Adherent cell lines can be shielded from shear forces by using porous carriers, and macroporous carriers are advantageous despite mass transfer limitations.…”

Section: Bioreactor Selectionmentioning

confidence: 99%

Concepts for the Production of Viruses and Viral Vectors in Cell Cultures

Grein¹,

Weidner²,

Czermak³

2017

New Insights Into Cell Culture Technology

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The industrial-scale manufacturing of viruses or virus-like particles in cell culture is necessary for gene therapy and the treatment of cancer with oncolytic viruses. Complex multistep processes are required in both cases, but the low virus titers in batch cultures and the temperature sensitivity of the virus particles limit the production scale. To meet commercial and regulatory requirements, each process must be scalable and reproducible and must yield high virus titers. These requirements are met by establishing a cell culture process that matches the properties of the virus/host-cell system and by using serum-free cell culture medium. This chapter focuses on two case studies to consider the different aspects of process design, such as the reactor configuration and operational mode: the continuous production of retroviral pseudotype vectors in a retroviral packaging cell line and the production of oncolytic measles virus vectors for cancer therapy.

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Section: Bioreactor Selectionmentioning

confidence: 99%

Concepts for the Production of Viruses and Viral Vectors in Cell Cultures

Grein¹,

Weidner²,

Czermak³

2017

New Insights Into Cell Culture Technology

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“…Three-phase systems, such as aerated microcarrier-based stirred-tank cultures, show the complex effects of the working volume, carrier concentration and aeration on the power input. Understanding the mixing characteristics of such a system is required to optimize hMSC cultivation [55]. Similarly, for hollow fiber or FBR systems, the flow rate must be defined according to the cell line, microcarrier and system volume [43].…”

Section: Cell Expansion In Bioreactorsmentioning

confidence: 99%

“…During cultivation, the stirrer speed was increased from 55 to 75 rpm and minimal aggregate formation was observed [46]. The mixing characteristics of this three-phase system showed a certain degree of inhomogeneity, but this was beneficial because the cells were allowed rest phases of low shear stress [55].…”

Section: Stirred Tank Reactormentioning

confidence: 99%

The Challenge of Human Mesenchymal Stromal Cell Expansion: Current and Prospective Answers

Elseberg¹,

Leber²,

Weidner³

et al. 2017

New Insights Into Cell Culture Technology

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In the field of cell therapy, allogenic human mesenchymal stromal cells (hMSCs) are often used in clinical trials, creating a demand for cell mass production using efficient dynamic bioreactor systems. As an advanced therapy medicinal product (ATMP), such cells should meet certain special requirements, including product specifications requiring a production process compatible with good manufacturing practice (GMP). The development of processes in which the cells are the product therefore remains a significant challenge. This chapter describes the requirements at different steps in the upstream and downstream phases of such dynamic processes. Potential solutions are presented and future prospects are discussed, including the selection of media and carriers for the strictly adherent growing cells, allowing efficient cell adhesion and detachment. Strategies for dynamic cultivation in bioreactors are described in detail for fixed-bed and stirred-tank reactors based on GMP requirements and the integration of process analytical technology (PAT). Following cell harvest, separation and purification, the formulation and storage of the product are also described. Finally, the chapter covers important cell quality characteristics necessary for the approval of ATMPs.

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“…One of the approaches to address this issue is to optimize configuration and geometry of stirred bioreactors and their impellers for maximum media mixing and minimum hydrodynamic shear stress. Numerous studies (Cioffi et al, ; Dusting, Sheridan, & Hourigan, ; Grein et al, ; Odeleye, Marsh, Osborne, Lye, & Micheletti, ; Santiago, de Campos Giordano, & Suazo, ; Sucosky, Osorio, Brown, & Neitzel, ) made improvements to a certain degree, yet they cannot overcome the fundamental limits imposed by the finite diffusion rate of gases and nutrients and the hydrodynamics. Another approach is to locally shield cells from the hydrodynamic shear stress.…”

Section: Introductionmentioning

confidence: 99%

“…Numerous studies (Cioffi et al, 2008;Dusting, Sheridan, & Hourigan, 2006;Grein et al, 2016;Odeleye, Marsh, Osborne, Lye, & Micheletti, 2014;Santiago, de Campos Giordano, & Suazo, 2011;Sucosky, Osorio, Brown, & Neitzel, 2004) made improvements to a certain degree, yet they cannot overcome the fundamental limits imposed by the finite diffusion rate of gases and nutrients and the hydrodynamics. Another approach is to locally shield cells from the hydrodynamic shear stress.…”

mentioning

confidence: 99%

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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Multiphase mixing characteristics in a microcarrier-based stirred tank bioreactor suitable for human mesenchymal stem cell expansion

Cited by 28 publications

References 37 publications

Concepts for the Production of Viruses and Viral Vectors in Cell Cultures

Concepts for the Production of Viruses and Viral Vectors in Cell Cultures

The Challenge of Human Mesenchymal Stromal Cell Expansion: Current and Prospective Answers

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

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