Considerations on the lactate consumption by CHO cells in the presence of galactose

Altamirano, Claudia; Illanes, Andrés; Becerra, Silvana; Cairó, Jordi J.; Gòdia, Francesc

doi:10.1016/j.jbiotec.2006.03.023

Cited by 160 publications

(153 citation statements)

References 24 publications

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“…Neither the apoptotic R nor the control cell lines exhibited sustained growth in the complete absence of glucose, that is, in the presence of lactate only (data not shown). Indeed, similar observations were made by Altamirano et al (2004Altamirano et al ( , 2006, where the authors showed that CHO cells were incapable of sustained growth in medium containing lactate as the sole carbon source. When both cell lines were cultured in the same custom-formulated medium containing ''high'' (60 mM) concentrations of glucose, EAX197 exhibited a higher VCD, viability and IVCC ( Figure 9A-C) than the control cell line especially at later times in the cell culture.…”

Section: Batch Culture In Presence Of High Glucosesupporting

confidence: 72%

Expression of anti‐apoptosis genes alters lactate metabolism of Chinese Hamster Ovary cells in culture

Dorai

Kyung

Ellis

et al. 2009

Biotech & Bioengineering

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ABSTRACT:In an effort to develop robust Chinese Hamster Ovary host cell lines, a variety of anti-apoptotic genes were over-expressed, either singly or in combination, followed by screening of transfectants for improved cell growth, extended longevity, reduced caspase 3/7 activity, and enhanced mitochondrial membrane potential (MMP). Two particular cell lines, one containing two anti-apoptotic genes, E1B-19K and Aven (EA167), and another containing three, E1B-19K, Aven, and a mutant of XIAP (EAX197), exhibited a reduction in caspase 3 activity of at least 60% and a 170% enhancement in mitochondrial membrane potential compared to controls when treated with staurosporine. In batch cell growth experiments, the peak viable cell densities and viabilities were higher resulting in a 186% increase in integrated viable cell densities. Analyses of metabolite utilization and formation of waste products indicated that the apoptotic resistant cell lines depleted all the lactate when grown in commercially available CD-CHO medium while significant levels (>1.8 g/L) accumulated in the host cell lines. When the lactate level was replenished daily in the apoptotic resistant cell lines, the cell lines consumed lactate and the culture longevity was extended up to four additional days compared to control cell lines. Furthermore, the anti-apoptosis cell lines also accumulated lower levels of ammonia. The ability of the apoptotic resistant cell lines to consume lactate was exploited by cultivating them in a ''high'' glucose medium containing 15 g/L (60 mM glucose) in which apoptotic resistant cell lines exhibited lower maximum lactate (1.8 g/L) compared to control cell lines which accumulated concentrations of lactate (2.2 g/L) that appeared to be deleterious for growth. The shaker flask titer of a therapeutic antibody product expressed in an apoptotic resistant cell line in ''high'' glucose medium reached 690 mg/ L compared to 390 mg/L for a cell line derived from a control host cell line. These results represent to our knowledge the first example in the literature in which manipulation of the apoptosis pathway has altered the nutrient consumption profile of mammalian cells in culture; findings that underscore the interdependence of the apoptotic cellular machinery and metabolism and provide greater flexibility to mammalian bioreactor process development.

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Section: Batch Culture In Presence Of High Glucosesupporting

confidence: 72%

Expression of anti‐apoptosis genes alters lactate metabolism of Chinese Hamster Ovary cells in culture

Dorai

Kyung

Ellis

et al. 2009

Biotech & Bioengineering

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“…Later during the culture, when the glucose concentration decreases, the fraction of the glucose implied in the TCA cycle increases. This result has been already observed by several authors (Altamirano et al 2006;Meuwly 2006). …”

Section: Analysis Of the Glucose Metabolismsupporting

confidence: 87%

Structured Model of VERO Cells Metabolism in a Fixed-Bed Bioreactor

Gelbgras¹,

Wylock²,

Drugmand³

et al. 2011

Chemical Product and Process Modeling

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In this work, a structured mathematical model of the VERvet Origin (VERO) cell metabolism in a fixed-bed bioreactor is developed. Mass balance equations for the extra-and intracellular species are written considering the main pathways of the animal cell metabolism. From the model equations, an undetermined set of equations relating the consumption or production velocities of the extracellular species and the velocities of the metabolic reactions can be developed. The monitoring of a reference cell culture enables to transform this undetermined set of equations in a determined one. The resolution of this determined set of equations enables to analyze and characterize the cell metabolism. From the model and the cell metabolism characterization, a monitoring tool is developed to simulate the time evolution of the species concentrations when the time evolution of the cell concentration is known. This simulation tool is used to simulate five experimental cell cultures. The comparison of the experimentally determined and computed time evolutions of the species concentrations enables to validate the developed monitoring tool. The validated monitoring tool can be used to optimize the operating conditions of the culture, as the medium feeding rate, the range of the species concentrations.

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“…Recently improved growth and t-PA productivity has been achieved by growing CHO cells in the presence of glutamate (6 mM) with 5 mM glucose and 20 mM galactose compared to a standard culture containing 6 mM glutamate and 20 mM glucose (Altamirano et al 2006). CHO initially consumed the glucose and then they continued growing on galactose in a second phase with the simultaneous consumption of the endogenous lactate and this resulted in improved culture longevity and reduced waste accumulation (Altamirano et al 2006). …”

Section: Media Formulation Approaches For Nutrient Based Cell Prolifementioning

confidence: 99%

Proliferation control strategies to improve productivity and survival during CHO based production culture

2007

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Chinese Hamster Ovary cells are the primary system for the production of recombinant proteins for therapeutic use. Protein productivity is directly proportional to viable biomass, viability and culture longevity of the producer cells and a number of approaches have been taken to optimise these parameters. Cell cycle arrest, particularly in G1 phase, typically using reduced temperature cultivation and nutritional control have been used to enhance productivity in production cultures by prolonging the production phase, but the mechanism by which these approaches work is still not fully understood. In this article, we analyse the public literature on proliferation control approaches as they apply to production cell lines with particular reference to what is known about the mechanisms behind each approach.

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Considerations on the lactate consumption by CHO cells in the presence of galactose

Cited by 160 publications

References 24 publications

Expression of anti‐apoptosis genes alters lactate metabolism of Chinese Hamster Ovary cells in culture

Expression of anti‐apoptosis genes alters lactate metabolism of Chinese Hamster Ovary cells in culture

Structured Model of VERO Cells Metabolism in a Fixed-Bed Bioreactor

Proliferation control strategies to improve productivity and survival during CHO based production culture

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