Simpler noninstrumented batch and semicontinuous cultures provide mammalian cell kinetic data comparable to continuous and perfusion cultures

Henry, Olivier; Kwok, Ezra; Piret, James M.

doi:10.1002/btpr.17

Cited by 37 publications

(25 citation statements)

References 45 publications

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“…However this latter option is sometimes adopted to reduce the labor burden. In systems absent of limitations and in growth phase, batch, semi-perfusion, semi-continuous, chemostat and perfusion systems exhibited comparable cell specific production and consumption rates given that the abundance of the nutrients is similar (Schmid et al 1992;Henry et al 2008).…”

Section: Screening Systemsmentioning

confidence: 90%

Perfusion Processes

Chotteau

2014

Cell Engineering

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The interest for perfusion is increasing nowadays. This new focus has emerged from a synergy of a demand for disposable equipment and the availability of robust cell separation device, as well as the need for higher flexibility and lower investment cost. The cell separation devices mostly used today are based on filtration, i.e. alternating flow filtration, tangential flow filtration, spin-filter, or acceleration/gravity, i.e. inclined settler, centrifuge, acoustic settler. This paper gives an introduction to the basic concepts of perfusion and its practical implementation. It reviews the actual cell separation devices and describes the approaches used in the field to develop and optimize the perfusion processes.

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Section: Screening Systemsmentioning

confidence: 90%

Perfusion Processes

Chotteau

2014

Cell Engineering

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“…nutrient concentrations), provided that pseudo-steady conditions can be achieved. Alternatively, steady-state operation may be experimentally approximated with the use of semi-continuous cultures [64], thereby alleviating the need for a complex small-scale chemostat setup and retaining the relative ease of batch cultivation.…”

Section: Discussionmentioning

confidence: 99%

Unraveling the metabolism of HEK-293 cells using lactate isotopomer analysis

Henry

Jolicœur

Kamen

2010

Bioprocess Biosyst Eng

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HEK-293 is the most extensively used human cell line for the production of viral vectors and is gaining increasing attention for the production of recombinant proteins by transient transfection. To further improve the metabolic characterization of this cell line, we have performed cultures using ¹³C-labeled substrates and measured the resulting mass isotopomer distributions in lactate by LC/MS. Simultaneous metabolite and isotopomer balancing allowed improvement and validation of the metabolic model and quantification of key intracellular pathways. We have determined the amounts of glucose carbon channeled through the PPP, incorporated into the TCA cycle for energy production and lipids biosynthesis, as well as the cytosolic and mitochondrial malic enzyme fluxes. Our analysis also revealed that glutamine did not significantly contribute to lactate formation. An improved and quantitative understanding of the central carbon metabolism is greatly needed to pursue the rational development of engineering approaches at both the cellular and process levels.

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“…For conducting the isotopic tracer studies, cells were inoculated at 2 × 10 5 cells mL −1 in basal medium supplemented with unlabeled glucose and glutamine at 37°C in a 5% CO 2 and 95% humid atmosphere. Following induction with 1 µg mL −1 of cumate at 48 h, the cultures were operated in semicontinuous mode, whereby a fixed culture volume was replaced by fresh medium on a daily basis so as to maintain pseudo‐constant cell and nutrient concentrations . Two sets of semicontinuous cultures were performed, one in which the medium was supplemented with 1 mM of glutamine, while the second set was supplemented with 5 mM.…”

Section: Methodsmentioning

confidence: 99%

“…For semicontinuous cultures, the cell specific rates were determined from pseudo‐steady state measurements and the daily medium exchange rate, as described in Ref. :

q_{s} = \frac{F}{V} \frac{(S_{f e e d} - S)}{X_{v}}

where F correspond to the number of volume exchanges per time unit.…”

Section: Methodsmentioning

confidence: 99%

Elucidating the effects of postinduction glutamine feeding on the growth and productivity of CHO cells

Sheikholeslami

Jolicœur

Henry

2014

Biotechnology Progress

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Inducible mammalian expression systems are increasingly being used for the production of valuable therapeutics. In such system, maximizing the product yield is achieved by carefully balancing the biomass concentration during the production phase and the specific productivity of the cells. These two factors are largely determined by the availability of nutrients and/or the presence of toxic waste metabolites in the culture environment. Glutamine is one of the most important components of cell culture medium, since this substrate is an important building block and source of energy for biomass and recombinant protein production. Its metabolism, however, ultimately leads to the formation of ammonia, a well known inhibitor of cellular growth and productivity. In this work, we show that nutrient feeding post-induction can greatly enhance the product yield by alleviating early limitations encountered in batch. Moreover, varying the amount of glutamine in the feed yielded two distinct culture behaviors post-induction; whereas excess glutamine allowed to reach greater cell concentrations, glutamine-limited fed-batch led to increased cell specific productivity. These two conditions also showed distinctive lactate metabolism. To further assess the physiological impact of glutamine levels on the cells, a comparative (13) C-metabolic flux analysis was conducted and a number of key intracellular fluxes were found to be affected by the amount of glutamine present in the feed during the production phase. Such information may provide useful clues for the identification of physiological markers of cell growth and productivity that could further guide the optimization of inducible expression systems.

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Simpler noninstrumented batch and semicontinuous cultures provide mammalian cell kinetic data comparable to continuous and perfusion cultures

Cited by 37 publications

References 45 publications

Perfusion Processes

Perfusion Processes

Unraveling the metabolism of HEK-293 cells using lactate isotopomer analysis

Elucidating the effects of postinduction glutamine feeding on the growth and productivity of CHO cells

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