A rotating bed system bioreactor enables cultivation of primary osteoblasts on well‐characterized sponceram® regarding structural and flow properties

Suck, Kirstin; Roeker, Stefanie; Diederichs, Solvig; Anton, Fabienne; Sanz-Herrera, José A.; Ochoa, Ignacio; Doblaré, M.; Scheper, Thomas; Griensven, Martijn van; Kasper, Cornelia

doi:10.1002/btpr.386

Cited by 13 publications

(7 citation statements)

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“…Furthermore, fluid flow within the culture vessel could provide mechanical stimulation of the cells [21, 22]. Perfusion bioreactors are widely used for in vitro cultivation of three-dimensional tissues [17-19] and for cell expansion under GMP conditions [20]. Several other studies have shown enhanced synthesis of matrix proteins in three dimensional cultures in perfusion bioreactor systems as compared to static culture systems [38-40].…”

Section: Discussionmentioning

confidence: 99%

“…Perfusion bioreactor systems are widely used for the cultivation of cells on scaffold materials for the in vitro fabrication of tissue engineered constructs [17-19]. Insufficient migration and growth of cells into the scaffold and insufficient nutrient transport, especially to the interior of the carrier material cultured under static conditions, can be overcome.…”

Section: Introductionmentioning

confidence: 99%

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Large Scale Expansion of Human Umbilical Cord Cells in a Rotating Bed System Bioreactor for Cardiovascular Tissue Engineering Applications

Reichardt¹,

Polchow²,

Shakibaei³

et al. 2013

TOBEJ

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Widespread use of human umbilical cord cells for cardiovascular tissue engineering requires production of large numbers of well-characterized cells under controlled conditions. In current research projects, the expansion of cells to be used to create a tissue construct is usually performed in static cell culture systems which are, however, often not satisfactory due to limitations in nutrient and oxygen supply. To overcome these limitations dynamic cell expansion in bioreactor systems under controllable conditions could be an important tool providing continuous perfusion for the generation of large numbers of viable pre-conditioned cells in a short time period. For this purpose cells derived from human umbilical cord arteries were expanded in a rotating bed system bioreactor for up to 9 days. For a comparative study, cells were cultivated under static conditions in standard culture devices.Our results demonstrated that the microenvironment in the perfusion bioreactor was more favorable than that of the standard cell culture flasks. Data suggested that cells in the bioreactor expanded 39 fold (38.7 ± 6.1 fold) in comparison to statically cultured cells (31.8 ± 3.0 fold). Large-scale production of cells in the bioreactor resulted in more than 3 x 108 cells from a single umbilical cord fragment within 9 days. Furthermore cell doubling time was lower in the bioreactor system and production of extracellular matrix components was higher. With this study, we present an appropriate method to expand human umbilical cord artery derived cells with high cellular proliferation rates in a well-defined bioreactor system under GMP conditions.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Large Scale Expansion of Human Umbilical Cord Cells in a Rotating Bed System Bioreactor for Cardiovascular Tissue Engineering Applications

Reichardt¹,

Polchow²,

Shakibaei³

et al. 2013

TOBEJ

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“…For bone tissue engineering, several approaches such as rotational reactors, perfusion reactors, or stretch and pressure chambers have been used to imitate the physiologic environment for bone formation and repair. 6,14,15,19,46–48 Besides the successful cultivation of bone substitutes in various other reactor types, perfusion culture is still the most common concept used for bone tissue engineering. 4 Several other bioreactor models have been already characterized by CFD modeling of internal flow and pressure distribution.…”

Section: Discussionmentioning

confidence: 99%

“…Experimental results are consistent with previous reports on the cultivation of tissue-engineered bone substitutes in perfusion bioreactors. 6,16,17,20,23,52 The problem relating to low yields of new tissue formation in a bioreactor may also be related to using a stiff matrix that can induce the early differentiation of stem cells, and the mono-cellular culture system which is not mimicked by nature in the same way. We propose that our system is suitable for studies of mechanical forces on cell proliferation and differentiation in a defined flow and pressure environment.…”

Section: Discussionmentioning

confidence: 99%

A Differential Pressure Laminar Flow Reactor Supports Osteogenic Differentiation and Extracellular Matrix Formation from Adipose Mesenchymal Stem Cells in a Macroporous Ceramic Scaffold

Weyand

Kasper

Israelowitz³

et al. 2012

BioResearch Open Access

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We present a laminar flow reactor for bone tissue engineering that was developed based on a computational fluid dynamics model. The bioreactor design permits a laminar flow field through its specific internal shape. An integrated bypass system that prevents pressure build-up through bypass openings for pressure release allows for a constant pressure environment during the changing of permeability values that are caused by cellular growth within a porous scaffold. A macroporous ceramic scaffold, composed of zirconium dioxide, was used as a test biomaterial that studies adipose stem cell behavior within a controlled three-dimensional (3D) flow and pressure environment. The topographic structure of the material provided a basis for stem cell proliferation and differentiation toward the osteogenic lineage. Dynamic culture conditions in the bioreactor supported cell viability during long-term culture and induced cell cluster formation and extra-cellular matrix deposition within the porous scaffold, though no complete closure of the pores with new-formed tissue was observed. We postulate that our system is suitable for studying fluid shear stress effects on stem cell proliferation and differentiation toward bone formation in tissue-engineered 3D constructs.

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“…Furthermore, an adequate supply with nutrients should be maintained. Currently, commercial and tailor‐made bioreactors used for bone TE are rotating‐wall vessels [55, 61, 73, 74], rotating beds [53, 75], spinner‐flask systems [55, 61, 73, 76, 77], and direct perfusion systems [61, 77–79], among others. In rotating‐wall and spinner‐flask bioreactors, sufficient transport of nutrients cannot be guaranteed into the inner part of scaffolds with clinically relevant sizes [55].…”

Section: Control Of Culture Conditions – Musculoskeletal Applicationsmentioning

confidence: 99%

Identification of genes encoding N‐glycan processing β‐N‐acetylglucosaminidases in Trichoplusia ni and Bombyx mori: Implications for glycoengineering of baculovirus expression systems

Geisler

Jarvis

2009

Biotechnology Progress

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Glycoproteins produced by non-engineered insects or insect cell lines characteristically bear truncated, paucimannose N-glycans in place of the complex N-glycans produced by mammalian cells. A key reason for this difference is the presence of a highly specific N-glycan processing β-N-acetylglucosaminidase in insect, but not in mammalian systems. Thus, reducing or abolishing this enzyme could enhance the ability of glycoengineered insects or insect cell lines to produce complex N-glycans. Of the three insect species routinely used for recombinant glycoprotein production, the processing β-N-acetylglucosaminidase gene has been isolated only from Spodoptera frugiperda. Thus, the purpose of this study was to isolate and characterize the genes encoding this important processing enzyme from the other two species, Bombyx mori and Trichoplusia ni. Bioinformatic analyses of putative processing β-N-acetylglucosaminidase genes isolated from these two species indicated that each encoded a product that was, indeed, more similar to processing β-N-acetylglucosaminidases than degradative or chitinolytic β-N-acetylglucosaminidases. In addition, over-expression of each of these genes induced an enzyme activity with the substrate specificity characteristic of processing, but not degradative or chitinolytic enzymes. Together, these results demonstrated that the processing β-N-acetylglucosaminidase genes had been successfully isolated from Trichoplusia ni and Bombyx mori. The identification of these genes has the potential to facilitate further glycoengineering of baculovirus-insect cell expression systems for the production of glycosylated proteins.

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A rotating bed system bioreactor enables cultivation of primary osteoblasts on well‐characterized sponceram® regarding structural and flow properties

Cited by 13 publications

References 50 publications

Large Scale Expansion of Human Umbilical Cord Cells in a Rotating Bed System Bioreactor for Cardiovascular Tissue Engineering Applications

Large Scale Expansion of Human Umbilical Cord Cells in a Rotating Bed System Bioreactor for Cardiovascular Tissue Engineering Applications

A Differential Pressure Laminar Flow Reactor Supports Osteogenic Differentiation and Extracellular Matrix Formation from Adipose Mesenchymal Stem Cells in a Macroporous Ceramic Scaffold

Identification of genes encoding N‐glycan processing β‐N‐acetylglucosaminidases in Trichoplusia ni and Bombyx mori: Implications for glycoengineering of baculovirus expression systems

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