Relationship between oxygen consumption rate and cellular activity of mammalian cells cultured in serum‐free media

Yamada, Koji; Furushou, Shigeki; Sugahara, Takuya; Shirahata, Sanetaka; Murakami, Hiroki

doi:10.1002/bit.260360715

Cited by 33 publications

(11 citation statements)

References 9 publications

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“…[1][2][3][4][5][6][7][8][9] Oxygen uptake data are an indicator of cell density and metabolic rates such as glucose consumption and online q O2 measurements have been used to design feeding strategies and to control bioreactor operation. [10][11][12] In addition, q O2 information is also necessary for bioreactor design and scale-up given the low solubility of oxygen and the high oxygen transfer requirements of high cell density perfusion and fed-batch cultures.…”

Section: Introductionmentioning

confidence: 99%

Estimating cell specific oxygen uptake and carbon dioxide production rates for mammalian cells in perfusion culture

Goudar

Piret

Konstantinov

2011

Biotechnology Progress

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We present robust methods for online estimation of cell specific oxygen uptake and carbon dioxide production rates (q(O2) and q(CO2), respectively) during perfusion cultivation of mammalian cells. Perfusion system gas and liquid phase mass balance expressions for oxygen and carbon dioxide were used to estimate q(O2), q(CO2) and the respiratory quotient (RQ) for Chinese hamster ovary (CHO) cells in perfusion culture over 12 steady states with varying dissolved oxygen (DO), pH, and temperature set points. Under standard conditions (DO = 50%, pH = 6.8, T = 36.5°C), q(O2) and q(CO2) ranges were 5.14-5.77 and 5.31-6.36 pmol/cell day, respectively, resulting in RQ values of 0.98-1.14. Changes to DO had a slight reducing effect on respiration rates with q(O2) and q(CO2) values of 4.64 and 5.47, respectively, at DO = 20% and 4.57 and 5.12 at DO = 100%. Respiration rates were lower at low pH with q(O2) and q(CO2) values of 4.07 and 4.15 pmol/cell day at pH = 6.6 and 4.98 and 5.36 pmol/cell day at pH = 7. Temperature also impacted respiration rates with respective q(O2) and q(CO2) values of 3.97 and 4.02 pmol/cell day at 30.5°C and 5.53 and 6.25 pmol/cell day at 37.5°C. Despite these changes in q(O2) and q(CO2) values, the RQ values in this study ranged from 0.98 to 1.23 suggesting that RQ was close to unity. Real-time q(O2) and q(CO2) estimates obtained using the approach presented in this study provide additional quantitative information on cell physiology both during bioprocess development and commercial biotherapeutic manufacturing.

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Section: Introductionmentioning

confidence: 99%

Estimating cell specific oxygen uptake and carbon dioxide production rates for mammalian cells in perfusion culture

Goudar

Piret

Konstantinov

2011

Biotechnology Progress

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“…Due to the static seeding process, at the outer surfaces of the DBM‐scaffold, where the oxygen imaging sensor measures, cell density is higher than in the center, where the control sensor is localized (Wendt et al, ). Consequently, the higher cell number leads to greater cell metabolism resulting in higher oxygen consumption (Santoro et al, ; Yamada et al, ). A further explanation could be the distinct properties of the needle‐type control sensor.…”

Section: Discussionmentioning

confidence: 99%

Section: Characterization Of Cell-seeding Strategies By Application Omentioning

confidence: 99%

“…Oxygen consumption of in vitro cultured 2D-3D constructs depends primarily on cell metabolic activity and proliferation (Guarino et al, 2004; Janssen et al, 2006;Santoro et al, 2011;Yamada et al, 1990). An increase in the number of cells leads to a decrease of the oxygen concentration (Santoro et al, 2011;Yamada et al, 1990) and its continuing decline causes apoptosis or necrosis (Volkmer et al, 2008(Volkmer et al, , 2010. In our study, hMSCs were harvested at low confluence (30-40%, population L) or high confluence (70-80%, population H) and seeded at two different densities (0.5 and 1 million cells) into DBM scaffolds, followed by the measurement of the oxygen consumption for 24 h. Our results show a highly significant difference in oxygen consumption between the two populations ( Fig.…”

Section: Characterization Of Cell-seeding Strategies By Application Omentioning

confidence: 99%

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Oxygen mapping: Probing a novel seeding strategy for bone tissue engineering

Westphal

Jedelhauser

Liebsch

et al. 2016

Biotech & Bioengineering

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Bone tissue engineering (BTE) utilizing biomaterial scaffolds and human mesenchymal stem cells (hMSCs) is a promising approach for the treatment of bone defects. The quality of engineered tissue is crucially affected by numerous parameters including cell density and the oxygen supply. In this study, a novel oxygen‐imaging sensor was introduced to monitor the oxygen distribution in three dimensional (3D) scaffolds in order to analyze a new cell‐seeding strategy. Immortalized hMSCs, pre‐cultured in a monolayer for 30–40% or 70–80% confluence, were used to seed demineralized bone matrix (DBM) scaffolds. Real‐time measurements of oxygen consumption in vitro were simultaneously performed by the novel planar sensor and a conventional needle‐type sensor over 24 h. Recorded oxygen maps of the novel planar sensor revealed that scaffolds, seeded with hMSCs harvested at lower densities (30–40% confluence), exhibited rapid exponential oxygen consumption profile. In contrast, harvesting cells at higher densities (70–80% confluence) resulted in a very slow, almost linear, oxygen decrease due to gradual achieving the stationary growth phase. In conclusion, it could be shown that not only the seeding density on a scaffold, but also the cell density at the time point of harvest is of major importance for BTE. The new cell seeding strategy of harvested MSCs at low density during its log phase could be a useful strategy for an early in vivo implantation of cell‐seeded scaffolds after a shorter in vitro culture period. Furthermore, the novel oxygen imaging sensor enables a continuous, two‐dimensional, quick and convenient to handle oxygen mapping for the development and optimization of tissue engineered scaffolds. Biotechnol. Bioeng. 2017;114: 894–902. © 2016 The Authors. Biotechnology and Bioengineering Published by Wiley Periodicals, Inc.

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“…Mammalian cells are being increasingly used for the production of therapeutic proteins because of their ability to properly fold and glycosylate these proteins. Robust control of dissolved oxygen concentration in mammalian cell bioreactors is important, as cellular growth and protein productivity have been known to be influenced by dissolved oxygen concentration (1)(2)(3)(4)(5)(6)(7). The mode of oxygen supply and characterization of oxygen mass transfer in the bioreactor are especially important given the low solubility of oxygen in cell culture medium (∼6 mg/L at 37°C).…”

Section: Introductionmentioning

confidence: 99%

Experimental and Theoretical Analysis of Tubular Membrane Aeration for Mammalian Cell Bioreactors

Goudar

Michaels

et al. 2008

Biotechnol Progress

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A combination of experimental and theoretical approaches was used to characterize the dynamics of oxygen transfer in a membrane-aerated bioreactor. Pressure profiles along the length of the membrane at varying entrance and exit pressures were determined by actual experimental measurements, unlike most previous studies that have relied solely on theoretical descriptions of the pressure profile in the tubing. The mass transfer coefficient, k(L)a, was also determined under these conditions and was found to be essentially independent of tubing exit pressure. Measurement of the tubing pressure profile coupled with estimation of k(L)a allowed for computation of the oxygen transfer rate (OTR) along the length of the tubing. A mathematical model that incorporated friction pressure loss and losses due to tubing bending was developed to describe the pressure and hence OTR characteristics of membrane-aerated systems. The applicability of the model was verified by testing it on experimentally measured pressure data, and in all cases the model accurately described experimental data. When tubing properties are known, the mathematical model presented in this study allows for a priori estimation of OTR profiles along the length of the tubing. This information is vital for optimal design and scale-up of membrane-aerated bioreactors for mammalian cell culture.

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Relationship between oxygen consumption rate and cellular activity of mammalian cells cultured in serum‐free media

Cited by 33 publications

References 9 publications

Estimating cell specific oxygen uptake and carbon dioxide production rates for mammalian cells in perfusion culture

Estimating cell specific oxygen uptake and carbon dioxide production rates for mammalian cells in perfusion culture

Oxygen mapping: Probing a novel seeding strategy for bone tissue engineering

Experimental and Theoretical Analysis of Tubular Membrane Aeration for Mammalian Cell Bioreactors

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